https://wiki.fkkt.uni-lj.si/api.php?action=feedcontributions&feedformat=atom&user=Roberta+Mulac 2026-07-01T08:18:51Z User contributions MediaWiki 1.45.3 https://wiki.fkkt.uni-lj.si/index.php?title=Cockroach_terminator&diff=14693 2018-12-17T17:52:29Z

<p>Roberta Mulac: </p> <hr /> <div>The SZU-China team won the golden medal for best manufacturing project and was the second runner up for the Grand prize. Web page of SZU-China team is: http://2018.igem.org/Team:SZU-China&lt;ref&gt;[1]&lt;/ref&gt;. <br /> ='''''Introduction'''''=<br /> Cockroaches are among the most common household pests, they harbor in damp and unsanitary places such as sewers, garbage disposals, kitchens, and bathroom, feed on human’s and pet’s food. Cockroaches have been implicated in the transmission of several pathogenic organisms such as E.coli and Salmonella enteritidis which can cause diarrhea, pneumonia and so on. &lt;ref name=[2]&gt;Mpuchane S, Matsheka IM, Gashe BA, Allotey J, Murindamombe G, Mrema N. Microbiological studies of cockroaches from three localities in Gaborone, Botswana. Afr J Food Nutr Sci. 2006;6:56–59&lt;/ref&gt;<br /> Currently, there are two ways to control the population of cockroaches, physical and chemical methods. However, a chemical method like foggers, boric acid, and gel bait may cause environmental toxicity and the development of resistance in cockroaches. A physical method such as “cockroach house”, only trap few cockroaches. Neither chemical nor physical method can’t achieve the goal of environment-friendly and efficient control. &lt;ref&gt;[3]&lt;/ref&gt;<br /> <br /> ='''''Chassis and parts'''''=<br /> The SZU-China team choose a kind of entomogenous fungi, Metarhizium anisopliae, as biological chassis. It is considered a safe and prospective choice for causing disease in insects only. &lt;ref&gt;[4]&lt;/ref&gt;<br /> Its infection process is divided into the following steps: <br /> <br /> 1. spore attachment <br /> <br /> 2. penetration through the body wall<br /> <br /> 3. colonization in vivo <br /> <br /> 4. killing the host<br /> <br /> 5. re-sporulation.<br /> <br /> It is a natural insecticide, and by genetically-enhancing it, we can make better use of it.<br /> But its lethality is still needed to be improved, which limits it's widespread used. They construct a system to enhance its virulence. Their system contains three parts: HsbA, BbChit, and MCL1. These three genes work sequentially during infecting cockroach, promote adhesion, penetration, and immune-avoidance respectively.<br /> HsbA- The HsbA from Beauveria bassiana (the entomopathogenic fungi, used as a biological insecticide) encodes a membrane surface hydrophobic protein A, located on the surface of fungus. This protein works like “glue” by forming a hydrophobic bond between spores and the waxy epicuticle of their host. &lt;ref&gt;[5]&lt;/ref&gt;<br /> <br /> BbChit- Cockroaches’ body wall is composed of protein, chitin, and lipids. BbChit, which comes from Beauveria bassiana, encodes chitinase which can hydrolize chitin to penetrate the host. Thereby, fungus can enter into hemolymph. &lt;ref&gt;[6]&lt;/ref&gt;<br /> <br /> MCL1-It encodes a collagen-like protein, which can combine with β-glucan. β-glucan is distributed on the cell wall surface of fungus and is the recognization site for insect hemocytes to combine and clear invader. MCL1 acts like putting an “invisible cloak” on the fungus, so that fungus can evade being recognized by the host immune system. Cockroaches have powerful immune system. If there is no effective response, their chassis won't be recognized and killed by host hemocytes. The MCL1 gene from Metarhizium Robertsii can encode collagen-like protein, which binds and masks β-1,3-glucan (an antigen that can be recognized by hemocytes), thus Metarhizium anisopliae can escape the immune response of cockroaches. &lt;ref&gt;[7]&lt;/ref&gt;<br /> ='''''Safety mechanism'''''=<br /> They designed a suicide switch for their system. It consists of tryptophan attenuator and MazF (venom protein). Tryptophan attenuator will be switched on in high Trp concentration, while it will be switched off on the contrary. To make MazF expressed in chassis, they added a Kozak sequence in front of these two parts. They create a high Tryptophan concentration environment while culturing the fungus, and average concentration of Trp inside cockroach hemolymph is about 0.15%. In those environments, the tryptophan attenuator forms a hairpin structure that stop transcription ahead of MazF, so that fungus can stay alive. If the fungus live in the low tryptophan environment , the suicide switch will be turned on and the MazF starts to express, which leads to death of fungus. &lt;ref&gt;[8]&lt;/ref&gt;<br /> ='''''Construction of plasmid vector'''''=<br /> They used the fungal plasmid pBC for their project, which can propagate in Metarhizium anisopliae. The vector contains the following parts:<br /> <br /> PgpD is the constitutive promoter of the glyceraldehyde-3-phosphate dehydrogenase gene (gpdA) from Aspergillus nidulans;<br /> TtrpC is a tryptophan terminator from Aspergillus nidulans;<br /> HsbA is encoded by the gene HsbA from Beauveria bassiana; <br /> BbChit is encoded by the gene Bbchit from Beauveria bassiana; <br /> MCL1_Metarhizium robertsii and <br /> Tryptophan-MazF-suicide switch, induces apoptosis in the absence of tryptophan.<br /> <br /> Inside the vectors, there are four gene pathways. PgpdA promoter starts the transcription of HsbA, Bbchit, MCL1, Ttyptophan-MazF. TtrpC terminator terminates the transcription. It is well to be reminded that the pathway, PgpdA-Ttyptophan-MazF-TtrpC has a special switch, which induces apoptosis in the absence of tryptophan.<br /> ='''''Results'''''=<br /> ==''Adhesion:HsbA''==<br /> In this part, a strong promoter, PgpdA allows the HsbA protein to be expressed without induction. In the HsbA macro verification protocol, they could compare whether there was any change of the position and number of spores in the observing area. The wild-type Metarhizium anisopliae groups had the adherence rate in average of 26.7%. The Metarhizium anisopliae HsbA transformant groups had the adherence rate in average of 97.7%. In conclusion, this result confirmed that Metarhizium anisopliae HsbA transformant certainly enhanced the capacity of adhesion.<br /> ==''Penetration:BbChit''==<br /> In order to verify the function of Bbchit, they improved Kan Zhuo's chitin transparent circle method for verification. They stained the czapek solid medium without chitin colloids in red with 0.1% Congo red dye solution and then inoculated wild-type and transformed Metarhizium.Then they compared the size of the colony and the transparent circle between wild type and transformant. The size measured is the diameter of the chitin transparent ring (R2) and colony(R1). The ratio of the diameters showed as R2/R1.The conclusion is that the chitinase activity of transformed Metarhizium anisopliae is enhanced by 1.3 times.<br /> ==''Immune avoidance:MCL1''==<br /> In this part, PgpdA is a strong promoter that allows MCL1 to be expressed without induction. Change of nodules formed of hemocytes in cockroach hemolymph after injecting hyphae homogenate. At the time point of 0.5h, 1h and 8h, the nodules caused by M.anasopliae is significantly higher than transformants´ which means immune-avoidance occurs in genetically enhanced M.anasopliae.<br /> ==''Suicide switch''==<br /> In order to confirm the limited concentration of tryptophan, they did a macro experiment. They put Metarhizium in an L-Tryptophan concentration gradient Petrie dishes from 0.05% to 0.14% with solid and liquid czapek. It was seen that the Metarhizium can stay alive at the L-Trp concentration of 0.09% or higher while it could not grow well or die at a lower concentration. They could also see that Metarhizium could not survive without L-Tryptophan.<br /> ='''''GreenGround trapbox'''''=<br /> Using all four parts they designed a trap box named GreenGround to bring their idea into real world. They mixed spores, banana powder and oil together to form emulsifiable powder, which is applied to non-woven fabrics for use. They developed it in three generations.<br /> ==''First generation''==<br /> At first, they used oil solution to contain Metarhizium anisopliae. One area was a hollow cube to contain baits for cockroaches. There were breathable holes on top of it to make sure the smell of the baits got out. There were four entrances which allowed the cockroaches to enter. When cockroaches got in, they would get in contact with Metarhizium anisopliae which would kill them. And the cockroaches could also get out and infect other cockroaches by contacting them.<br /> After the human practice in Chongqing, they knew that cockroaches will be likely to climb gentle slope instead of entrances it.<br /> ==''Second generation''==<br /> They decided to use Metarhizium anisopliae emulsifiable powder on nonwoven fabric instead of oil solution. This time they mixed the baits with Metarhizium anisopliae emulsifiable powder and brush them on nonwoven fabric. They made a box which formed a cleft for the cockroaches to climb in through the gentle slope. In the middle were 5 magnet cubes, which could adjust the height of the cleft according to the actual size of the cockroach. They found out that magnets are not stable enough and that acrylic has its own limitations.<br /> ==''Final generation''==<br /> After they found acrylic can no longer fit their needs, they change the material into polypropylene. They also mix baits with Metarhizium anisopliae emulsifiable powder on polypropylene surface. They screw the lid down through the cylinder to form cleft which is adjustable. The cockroaches can climb through the slope and get in contact with Metarhizium anisopliae which will lead to death.<br /> ='''''Conclusion'''''=<br /> GreenGround trap box is safe and easy to use. It does not cause environmental toxicity and the development of resistance in cockroaches. Their improved M. anisopliae has better adhesion rate, is more successful in penetrating the host and can avoid immune system of the host, comparing to WT M. anisopliae.<br /> They wanted to make their product avalliable for the public since lots of poor neighborhoods in China have problem with cockroaches and other insects. They suceed in that by improving they trap box to maximize its function and to make it cheap. They also put their GreenGround box on popular app so they can get a feedback from public.<br /> ='''''Literature'''''=<br /> [1] http://2018.igem.org/Team:SZU-China<br /> <br /> [2] Mpuchane S, Matsheka IM, Gashe BA, Allotey J, Murindamombe G, Mrema N. Microbiological studies of cockroaches from three localities in Gaborone, Botswana. Afr J Food Nutr Sci. 2006;6:56–59<br /> <br /> [3] Wu X, Appel AG.J Econ Entomol. 2017 Jun 1;110(3):1203-1209<br /> <br /> [4] Wang C, Lü Dingding, Li Lin. Study on pathogenicity and degradation mechanism of entomogenous fungi [C]// Chinese Society of Fungal Sciences Academic Symposium. 2008<br /> <br /> [5] Ye Zhang, Zhongren Lei, Haihong Wang, Jiqing Zhan. Prokaryotic expression and immunolocalization Beauveria bassiana HsbA protein [J] Chinese Agricultural Sciences, 2013,46 (21): 4534-4541<br /> <br /> [6] Liu Zhihui, Chen Shouwen, Guo Zhihong, et al. Correlation between extracellular protease and chitinase activity of Beauveria bassiana and virulence to Asian corn borer[J]. Journal of Huazhong Agricultural University, 2005, 24(4) :364-368<br /> <br /> [7] Wang C, St Leger R J. A collagenous protective coat enables Metarhizium anisopliae to evade insect immune responses.[J]. Proceedings of the National Academy of Sciences of the United States of America, 2006, 103(17):6647-6652<br /> <br /> [8] Sowa S M, Keeley L L. Free amino acids in the hemolymph of the cockroach, Blaberus discoidalis[J]. Comparative Biochemistry &amp; Physiology Part A Physiology, 1996, 113(2):131</div>

Roberta Mulac https://wiki.fkkt.uni-lj.si/index.php?title=Cockroach_terminator&diff=14690 2018-12-17T17:38:55Z

<p>Roberta Mulac: </p> <hr /> <div>The SZU-China team won the golden medal for best manufacturing project and was the second runner up for the Grand prize. Web page of SZU-China team is: http://2018.igem.org/Team:SZU-China&lt;ref&gt;[1]&lt;/ref&gt;. <br /> ='''''Introduction'''''=<br /> Cockroaches are among the most common household pests, they harbor in damp and unsanitary places such as sewers, garbage disposals, kitchens, and bathroom, feed on human’s and pet’s food. Cockroaches have been implicated in the transmission of several pathogenic organisms such as E.coli and Salmonella enteritidis which can cause diarrhea, pneumonia and so on. &lt;ref name=[2]&gt;Mpuchane S, Matsheka IM, Gashe BA, Allotey J, Murindamombe G, Mrema N. Microbiological studies of cockroaches from three localities in Gaborone, Botswana. Afr J Food Nutr Sci. 2006;6:56–59&lt;/ref&gt;<br /> Currently, there are two ways to control the population of cockroaches, physical and chemical methods. However, a chemical method like foggers, boric acid, and gel bait may cause environmental toxicity and the development of resistance in cockroaches. A physical method such as “cockroach house”, only trap few cockroaches. Neither chemical nor physical method can’t achieve the goal of environment-friendly and efficient control. [3]<br /> <br /> ='''''Chassis and parts'''''=<br /> The SZU-China team choose a kind of entomogenous fungi, Metarhizium anisopliae, as biological chassis. It is considered a safe and prospective choice for causing disease in insects only. [4]<br /> Its infection process is divided into the following steps: <br /> <br /> 1. spore attachment <br /> <br /> 2. penetration through the body wall<br /> <br /> 3. colonization in vivo <br /> <br /> 4. killing the host<br /> <br /> 5. re-sporulation.<br /> <br /> It is a natural insecticide, and by genetically-enhancing it, we can make better use of it.<br /> But its lethality is still needed to be improved, which limits it's widespread used. They construct a system to enhance its virulence. Their system contains three parts: HsbA, BbChit, and MCL1. These three genes work sequentially during infecting cockroach, promote adhesion, penetration, and immune-avoidance respectively.<br /> HsbA- The HsbA from Beauveria bassiana (the entomopathogenic fungi, used as a biological insecticide) encodes a membrane surface hydrophobic protein A, located on the surface of fungus. This protein works like “glue” by forming a hydrophobic bond between spores and the waxy epicuticle of their host. [5]<br /> BbChit- Cockroaches’ body wall is composed of protein, chitin, and lipids. BbChit, which comes from Beauveria bassiana, encodes chitinase which can hydrolize chitin to penetrate the host. Thereby, fungus can enter into hemolymph. [6]<br /> MCL1-It encodes a collagen-like protein, which can combine with β-glucan. β-glucan is distributed on the cell wall surface of fungus and is the recognization site for insect hemocytes to combine and clear invader. MCL1 acts like putting an “invisible cloak” on the fungus, so that fungus can evade being recognized by the host immune system. Cockroaches have powerful immune system. If there is no effective response, their chassis won't be recognized and killed by host hemocytes. The MCL1 gene from Metarhizium Robertsii can encode collagen-like protein, which binds and masks β-1,3-glucan (an antigen that can be recognized by hemocytes), thus Metarhizium anisopliae can escape the immune response of cockroaches. [7]<br /> ='''''Safety mechanism'''''=<br /> They designed a suicide switch for their system. It consists of tryptophan attenuator and MazF (venom protein). Tryptophan attenuator will be switched on in high Trp concentration, while it will be switched off on the contrary. To make MazF expressed in chassis, they added a Kozak sequence in front of these two parts. They create a high Tryptophan concentration environment while culturing the fungus, and average concentration of Trp inside cockroach hemolymph is about 0.15%. In those environments, the tryptophan attenuator forms a hairpin structure that stop transcription ahead of MazF, so that fungus can stay alive. If the fungus live in the low tryptophan environment , the suicide switch will be turned on and the MazF starts to express, which leads to death of fungus. [8]<br /> ='''''Construction of plasmid vector'''''=<br /> They used the fungal plasmid pBC for their project, which can propagate in Metarhizium anisopliae. The vector contains the following parts:<br /> <br /> PgpD is the constitutive promoter of the glyceraldehyde-3-phosphate dehydrogenase gene (gpdA) from Aspergillus nidulans;<br /> TtrpC is a tryptophan terminator from Aspergillus nidulans;<br /> HsbA is encoded by the gene HsbA from Beauveria bassiana; <br /> BbChit is encoded by the gene Bbchit from Beauveria bassiana; <br /> MCL1_Metarhizium robertsii and <br /> Tryptophan-MazF-suicide switch, induces apoptosis in the absence of tryptophan.<br /> <br /> Inside the vectors, there are four gene pathways. PgpdA promoter starts the transcription of HsbA, Bbchit, MCL1, Ttyptophan-MazF. TtrpC terminator terminates the transcription. It is well to be reminded that the pathway, PgpdA-Ttyptophan-MazF-TtrpC has a special switch, which induces apoptosis in the absence of tryptophan.<br /> ='''''Results'''''=<br /> ==''Adhesion:HsbA''==<br /> In this part, a strong promoter, PgpdA allows the HsbA protein to be expressed without induction. In the HsbA macro verification protocol, they could compare whether there was any change of the position and number of spores in the observing area. The wild-type Metarhizium anisopliae groups had the adherence rate in average of 26.7%. The Metarhizium anisopliae HsbA transformant groups had the adherence rate in average of 97.7%. In conclusion, this result confirmed that Metarhizium anisopliae HsbA transformant certainly enhanced the capacity of adhesion.<br /> ==''Penetration:BbChit''==<br /> In order to verify the function of Bbchit, they improved Kan Zhuo's chitin transparent circle method for verification. They stained the czapek solid medium without chitin colloids in red with 0.1% Congo red dye solution and then inoculated wild-type and transformed Metarhizium.Then they compared the size of the colony and the transparent circle between wild type and transformant. The size measured is the diameter of the chitin transparent ring (R2) and colony(R1). The ratio of the diameters showed as R2/R1.The conclusion is that the chitinase activity of transformed Metarhizium anisopliae is enhanced by 1.3 times.<br /> ==''Immune avoidance:MCL1''==<br /> In this part, PgpdA is a strong promoter that allows MCL1 to be expressed without induction. Change of nodules formed of hemocytes in cockroach hemolymph after injecting hyphae homogenate. At the time point of 0.5h, 1h and 8h, the nodules caused by M.anasopliae is significantly higher than transformants´ which means immune-avoidance occurs in genetically enhanced M.anasopliae.<br /> ==''Suicide switch''==<br /> In order to confirm the limited concentration of tryptophan, they did a macro experiment. They put Metarhizium in an L-Tryptophan concentration gradient Petrie dishes from 0.05% to 0.14% with solid and liquid czapek. It was seen that the Metarhizium can stay alive at the L-Trp concentration of 0.09% or higher while it could not grow well or die at a lower concentration. They could also see that Metarhizium could not survive without L-Tryptophan.<br /> ='''''GreenGround trapbox'''''=<br /> Using all four parts they designed a trap box named GreenGround to bring their idea into real world. They mixed spores, banana powder and oil together to form emulsifiable powder, which is applied to non-woven fabrics for use. They developed it in three generations.<br /> ==''First generation''==<br /> At first, they used oil solution to contain Metarhizium anisopliae. One area was a hollow cube to contain baits for cockroaches. There were breathable holes on top of it to make sure the smell of the baits got out. There were four entrances which allowed the cockroaches to enter. When cockroaches got in, they would get in contact with Metarhizium anisopliae which would kill them. And the cockroaches could also get out and infect other cockroaches by contacting them.<br /> After the human practice in Chongqing, they knew that cockroaches will be likely to climb gentle slope instead of entrances it.<br /> ==''Second generation''==<br /> They decided to use Metarhizium anisopliae emulsifiable powder on nonwoven fabric instead of oil solution. This time they mixed the baits with Metarhizium anisopliae emulsifiable powder and brush them on nonwoven fabric. They made a box which formed a cleft for the cockroaches to climb in through the gentle slope. In the middle were 5 magnet cubes, which could adjust the height of the cleft according to the actual size of the cockroach. They found out that magnets are not stable enough and that acrylic has its own limitations.<br /> ==''Final generation''==<br /> After they found acrylic can no longer fit their needs, they change the material into polypropylene. They also mix baits with Metarhizium anisopliae emulsifiable powder on polypropylene surface. They screw the lid down through the cylinder to form cleft which is adjustable. The cockroaches can climb through the slope and get in contact with Metarhizium anisopliae which will lead to death.<br /> ='''''Conclusion'''''=<br /> GreenGround trap box is safe and easy to use. It does not cause environmental toxicity and the development of resistance in cockroaches. Their improved M. anisopliae has better adhesion rate, is more successful in penetrating the host and can avoid immune system of the host, comparing to WT M. anisopliae.<br /> They wanted to make their product avalliable for the public since lots of poor neighborhoods in China have problem with cockroaches and other insects. They suceed in that by improving they trap box to maximize its function and to make it cheap. They also put their GreenGround box on popular app so they can get a feedback from public.<br /> ='''''Literature'''''=<br /> [1] http://2018.igem.org/Team:SZU-China<br /> <br /> [2] Mpuchane S, Matsheka IM, Gashe BA, Allotey J, Murindamombe G, Mrema N. Microbiological studies of cockroaches from three localities in Gaborone, Botswana. Afr J Food Nutr Sci. 2006;6:56–59<br /> <br /> [3] Wu X, Appel AG.J Econ Entomol. 2017 Jun 1;110(3):1203-1209<br /> <br /> [4] Wang C, Lü Dingding, Li Lin. Study on pathogenicity and degradation mechanism of entomogenous fungi [C]// Chinese Society of Fungal Sciences Academic Symposium. 2008<br /> <br /> [5] Ye Zhang, Zhongren Lei, Haihong Wang, Jiqing Zhan. Prokaryotic expression and immunolocalization Beauveria bassiana HsbA protein [J] Chinese Agricultural Sciences, 2013,46 (21): 4534-4541<br /> <br /> [6] Liu Zhihui, Chen Shouwen, Guo Zhihong, et al. Correlation between extracellular protease and chitinase activity of Beauveria bassiana and virulence to Asian corn borer[J]. Journal of Huazhong Agricultural University, 2005, 24(4) :364-368<br /> <br /> [7] Wang C, St Leger R J. A collagenous protective coat enables Metarhizium anisopliae to evade insect immune responses.[J]. Proceedings of the National Academy of Sciences of the United States of America, 2006, 103(17):6647-6652<br /> <br /> [8] Sowa S M, Keeley L L. Free amino acids in the hemolymph of the cockroach, Blaberus discoidalis[J]. Comparative Biochemistry &amp; Physiology Part A Physiology, 1996, 113(2):131</div>

Roberta Mulac https://wiki.fkkt.uni-lj.si/index.php?title=Cockroach_terminator&diff=14689 2018-12-17T17:38:03Z

<p>Roberta Mulac: /* Introduction */</p> <hr /> <div>The SZU-China team won the golden medal for best manufacturing project and was the second runner up for the Grand prize. Web page of SZU-China team is: http://2018.igem.org/Team:SZU-China. <br /> ='''''Introduction'''''=<br /> Cockroaches are among the most common household pests, they harbor in damp and unsanitary places such as sewers, garbage disposals, kitchens, and bathroom, feed on human’s and pet’s food. Cockroaches have been implicated in the transmission of several pathogenic organisms such as E.coli and Salmonella enteritidis which can cause diarrhea, pneumonia and so on. &lt;ref name=[2]&gt;Mpuchane S, Matsheka IM, Gashe BA, Allotey J, Murindamombe G, Mrema N. Microbiological studies of cockroaches from three localities in Gaborone, Botswana. Afr J Food Nutr Sci. 2006;6:56–59&lt;/ref&gt;<br /> Currently, there are two ways to control the population of cockroaches, physical and chemical methods. However, a chemical method like foggers, boric acid, and gel bait may cause environmental toxicity and the development of resistance in cockroaches. A physical method such as “cockroach house”, only trap few cockroaches. Neither chemical nor physical method can’t achieve the goal of environment-friendly and efficient control. [3]<br /> <br /> ='''''Chassis and parts'''''=<br /> The SZU-China team choose a kind of entomogenous fungi, Metarhizium anisopliae, as biological chassis. It is considered a safe and prospective choice for causing disease in insects only. [4]<br /> Its infection process is divided into the following steps: <br /> <br /> 1. spore attachment <br /> <br /> 2. penetration through the body wall<br /> <br /> 3. colonization in vivo <br /> <br /> 4. killing the host<br /> <br /> 5. re-sporulation.<br /> <br /> It is a natural insecticide, and by genetically-enhancing it, we can make better use of it.<br /> But its lethality is still needed to be improved, which limits it's widespread used. They construct a system to enhance its virulence. Their system contains three parts: HsbA, BbChit, and MCL1. These three genes work sequentially during infecting cockroach, promote adhesion, penetration, and immune-avoidance respectively.<br /> HsbA- The HsbA from Beauveria bassiana (the entomopathogenic fungi, used as a biological insecticide) encodes a membrane surface hydrophobic protein A, located on the surface of fungus. This protein works like “glue” by forming a hydrophobic bond between spores and the waxy epicuticle of their host. [5]<br /> BbChit- Cockroaches’ body wall is composed of protein, chitin, and lipids. BbChit, which comes from Beauveria bassiana, encodes chitinase which can hydrolize chitin to penetrate the host. Thereby, fungus can enter into hemolymph. [6]<br /> MCL1-It encodes a collagen-like protein, which can combine with β-glucan. β-glucan is distributed on the cell wall surface of fungus and is the recognization site for insect hemocytes to combine and clear invader. MCL1 acts like putting an “invisible cloak” on the fungus, so that fungus can evade being recognized by the host immune system. Cockroaches have powerful immune system. If there is no effective response, their chassis won't be recognized and killed by host hemocytes. The MCL1 gene from Metarhizium Robertsii can encode collagen-like protein, which binds and masks β-1,3-glucan (an antigen that can be recognized by hemocytes), thus Metarhizium anisopliae can escape the immune response of cockroaches. [7]<br /> ='''''Safety mechanism'''''=<br /> They designed a suicide switch for their system. It consists of tryptophan attenuator and MazF (venom protein). Tryptophan attenuator will be switched on in high Trp concentration, while it will be switched off on the contrary. To make MazF expressed in chassis, they added a Kozak sequence in front of these two parts. They create a high Tryptophan concentration environment while culturing the fungus, and average concentration of Trp inside cockroach hemolymph is about 0.15%. In those environments, the tryptophan attenuator forms a hairpin structure that stop transcription ahead of MazF, so that fungus can stay alive. If the fungus live in the low tryptophan environment , the suicide switch will be turned on and the MazF starts to express, which leads to death of fungus. [8]<br /> ='''''Construction of plasmid vector'''''=<br /> They used the fungal plasmid pBC for their project, which can propagate in Metarhizium anisopliae. The vector contains the following parts:<br /> <br /> PgpD is the constitutive promoter of the glyceraldehyde-3-phosphate dehydrogenase gene (gpdA) from Aspergillus nidulans;<br /> TtrpC is a tryptophan terminator from Aspergillus nidulans;<br /> HsbA is encoded by the gene HsbA from Beauveria bassiana; <br /> BbChit is encoded by the gene Bbchit from Beauveria bassiana; <br /> MCL1_Metarhizium robertsii and <br /> Tryptophan-MazF-suicide switch, induces apoptosis in the absence of tryptophan.<br /> <br /> Inside the vectors, there are four gene pathways. PgpdA promoter starts the transcription of HsbA, Bbchit, MCL1, Ttyptophan-MazF. TtrpC terminator terminates the transcription. It is well to be reminded that the pathway, PgpdA-Ttyptophan-MazF-TtrpC has a special switch, which induces apoptosis in the absence of tryptophan.<br /> ='''''Results'''''=<br /> ==''Adhesion:HsbA''==<br /> In this part, a strong promoter, PgpdA allows the HsbA protein to be expressed without induction. In the HsbA macro verification protocol, they could compare whether there was any change of the position and number of spores in the observing area. The wild-type Metarhizium anisopliae groups had the adherence rate in average of 26.7%. The Metarhizium anisopliae HsbA transformant groups had the adherence rate in average of 97.7%. In conclusion, this result confirmed that Metarhizium anisopliae HsbA transformant certainly enhanced the capacity of adhesion.<br /> ==''Penetration:BbChit''==<br /> In order to verify the function of Bbchit, they improved Kan Zhuo's chitin transparent circle method for verification. They stained the czapek solid medium without chitin colloids in red with 0.1% Congo red dye solution and then inoculated wild-type and transformed Metarhizium.Then they compared the size of the colony and the transparent circle between wild type and transformant. The size measured is the diameter of the chitin transparent ring (R2) and colony(R1). The ratio of the diameters showed as R2/R1.The conclusion is that the chitinase activity of transformed Metarhizium anisopliae is enhanced by 1.3 times.<br /> ==''Immune avoidance:MCL1''==<br /> In this part, PgpdA is a strong promoter that allows MCL1 to be expressed without induction. Change of nodules formed of hemocytes in cockroach hemolymph after injecting hyphae homogenate. At the time point of 0.5h, 1h and 8h, the nodules caused by M.anasopliae is significantly higher than transformants´ which means immune-avoidance occurs in genetically enhanced M.anasopliae.<br /> ==''Suicide switch''==<br /> In order to confirm the limited concentration of tryptophan, they did a macro experiment. They put Metarhizium in an L-Tryptophan concentration gradient Petrie dishes from 0.05% to 0.14% with solid and liquid czapek. It was seen that the Metarhizium can stay alive at the L-Trp concentration of 0.09% or higher while it could not grow well or die at a lower concentration. They could also see that Metarhizium could not survive without L-Tryptophan.<br /> ='''''GreenGround trapbox'''''=<br /> Using all four parts they designed a trap box named GreenGround to bring their idea into real world. They mixed spores, banana powder and oil together to form emulsifiable powder, which is applied to non-woven fabrics for use. They developed it in three generations.<br /> ==''First generation''==<br /> At first, they used oil solution to contain Metarhizium anisopliae. One area was a hollow cube to contain baits for cockroaches. There were breathable holes on top of it to make sure the smell of the baits got out. There were four entrances which allowed the cockroaches to enter. When cockroaches got in, they would get in contact with Metarhizium anisopliae which would kill them. And the cockroaches could also get out and infect other cockroaches by contacting them.<br /> After the human practice in Chongqing, they knew that cockroaches will be likely to climb gentle slope instead of entrances it.<br /> ==''Second generation''==<br /> They decided to use Metarhizium anisopliae emulsifiable powder on nonwoven fabric instead of oil solution. This time they mixed the baits with Metarhizium anisopliae emulsifiable powder and brush them on nonwoven fabric. They made a box which formed a cleft for the cockroaches to climb in through the gentle slope. In the middle were 5 magnet cubes, which could adjust the height of the cleft according to the actual size of the cockroach. They found out that magnets are not stable enough and that acrylic has its own limitations.<br /> ==''Final generation''==<br /> After they found acrylic can no longer fit their needs, they change the material into polypropylene. They also mix baits with Metarhizium anisopliae emulsifiable powder on polypropylene surface. They screw the lid down through the cylinder to form cleft which is adjustable. The cockroaches can climb through the slope and get in contact with Metarhizium anisopliae which will lead to death.<br /> ='''''Conclusion'''''=<br /> GreenGround trap box is safe and easy to use. It does not cause environmental toxicity and the development of resistance in cockroaches. Their improved M. anisopliae has better adhesion rate, is more successful in penetrating the host and can avoid immune system of the host, comparing to WT M. anisopliae.<br /> They wanted to make their product avalliable for the public since lots of poor neighborhoods in China have problem with cockroaches and other insects. They suceed in that by improving they trap box to maximize its function and to make it cheap. They also put their GreenGround box on popular app so they can get a feedback from public.<br /> ='''''Literature'''''=<br /> [1] http://2018.igem.org/Team:SZU-China<br /> <br /> [2] Mpuchane S, Matsheka IM, Gashe BA, Allotey J, Murindamombe G, Mrema N. Microbiological studies of cockroaches from three localities in Gaborone, Botswana. Afr J Food Nutr Sci. 2006;6:56–59<br /> <br /> [3] Wu X, Appel AG.J Econ Entomol. 2017 Jun 1;110(3):1203-1209<br /> <br /> [4] Wang C, Lü Dingding, Li Lin. Study on pathogenicity and degradation mechanism of entomogenous fungi [C]// Chinese Society of Fungal Sciences Academic Symposium. 2008<br /> <br /> [5] Ye Zhang, Zhongren Lei, Haihong Wang, Jiqing Zhan. Prokaryotic expression and immunolocalization Beauveria bassiana HsbA protein [J] Chinese Agricultural Sciences, 2013,46 (21): 4534-4541<br /> <br /> [6] Liu Zhihui, Chen Shouwen, Guo Zhihong, et al. Correlation between extracellular protease and chitinase activity of Beauveria bassiana and virulence to Asian corn borer[J]. Journal of Huazhong Agricultural University, 2005, 24(4) :364-368<br /> <br /> [7] Wang C, St Leger R J. A collagenous protective coat enables Metarhizium anisopliae to evade insect immune responses.[J]. Proceedings of the National Academy of Sciences of the United States of America, 2006, 103(17):6647-6652<br /> <br /> [8] Sowa S M, Keeley L L. Free amino acids in the hemolymph of the cockroach, Blaberus discoidalis[J]. Comparative Biochemistry &amp; Physiology Part A Physiology, 1996, 113(2):131</div>

Roberta Mulac https://wiki.fkkt.uni-lj.si/index.php?title=Cockroach_terminator&diff=14686 2018-12-17T17:32:53Z

<p>Roberta Mulac: </p> <hr /> <div>The SZU-China team won the golden medal for best manufacturing project and was the second runner up for the Grand prize. Web page of SZU-China team is: http://2018.igem.org/Team:SZU-China. <br /> ='''''Introduction'''''=<br /> Cockroaches are among the most common household pests, they harbor in damp and unsanitary places such as sewers, garbage disposals, kitchens, and bathroom, feed on human’s and pet’s food. Cockroaches have been implicated in the transmission of several pathogenic organisms such as E.coli and Salmonella enteritidis which can cause diarrhea, pneumonia and so on. [2]<br /> Currently, there are two ways to control the population of cockroaches, physical and chemical methods. However, a chemical method like foggers, boric acid, and gel bait may cause environmental toxicity and the development of resistance in cockroaches. A physical method such as “cockroach house”, only trap few cockroaches. Neither chemical nor physical method can’t achieve the goal of environment-friendly and efficient control. [3]<br /> ='''''Chassis and parts'''''=<br /> The SZU-China team choose a kind of entomogenous fungi, Metarhizium anisopliae, as biological chassis. It is considered a safe and prospective choice for causing disease in insects only. [4]<br /> Its infection process is divided into the following steps: <br /> <br /> 1. spore attachment <br /> <br /> 2. penetration through the body wall<br /> <br /> 3. colonization in vivo <br /> <br /> 4. killing the host<br /> <br /> 5. re-sporulation.<br /> <br /> It is a natural insecticide, and by genetically-enhancing it, we can make better use of it.<br /> But its lethality is still needed to be improved, which limits it's widespread used. They construct a system to enhance its virulence. Their system contains three parts: HsbA, BbChit, and MCL1. These three genes work sequentially during infecting cockroach, promote adhesion, penetration, and immune-avoidance respectively.<br /> HsbA- The HsbA from Beauveria bassiana (the entomopathogenic fungi, used as a biological insecticide) encodes a membrane surface hydrophobic protein A, located on the surface of fungus. This protein works like “glue” by forming a hydrophobic bond between spores and the waxy epicuticle of their host. [5]<br /> BbChit- Cockroaches’ body wall is composed of protein, chitin, and lipids. BbChit, which comes from Beauveria bassiana, encodes chitinase which can hydrolize chitin to penetrate the host. Thereby, fungus can enter into hemolymph. [6]<br /> MCL1-It encodes a collagen-like protein, which can combine with β-glucan. β-glucan is distributed on the cell wall surface of fungus and is the recognization site for insect hemocytes to combine and clear invader. MCL1 acts like putting an “invisible cloak” on the fungus, so that fungus can evade being recognized by the host immune system. Cockroaches have powerful immune system. If there is no effective response, their chassis won't be recognized and killed by host hemocytes. The MCL1 gene from Metarhizium Robertsii can encode collagen-like protein, which binds and masks β-1,3-glucan (an antigen that can be recognized by hemocytes), thus Metarhizium anisopliae can escape the immune response of cockroaches. [7]<br /> ='''''Safety mechanism'''''=<br /> They designed a suicide switch for their system. It consists of tryptophan attenuator and MazF (venom protein). Tryptophan attenuator will be switched on in high Trp concentration, while it will be switched off on the contrary. To make MazF expressed in chassis, they added a Kozak sequence in front of these two parts. They create a high Tryptophan concentration environment while culturing the fungus, and average concentration of Trp inside cockroach hemolymph is about 0.15%. In those environments, the tryptophan attenuator forms a hairpin structure that stop transcription ahead of MazF, so that fungus can stay alive. If the fungus live in the low tryptophan environment , the suicide switch will be turned on and the MazF starts to express, which leads to death of fungus. [8]<br /> ='''''Construction of plasmid vector'''''=<br /> They used the fungal plasmid pBC for their project, which can propagate in Metarhizium anisopliae. The vector contains the following parts:<br /> <br /> PgpD is the constitutive promoter of the glyceraldehyde-3-phosphate dehydrogenase gene (gpdA) from Aspergillus nidulans;<br /> TtrpC is a tryptophan terminator from Aspergillus nidulans;<br /> HsbA is encoded by the gene HsbA from Beauveria bassiana; <br /> BbChit is encoded by the gene Bbchit from Beauveria bassiana; <br /> MCL1_Metarhizium robertsii and <br /> Tryptophan-MazF-suicide switch, induces apoptosis in the absence of tryptophan.<br /> <br /> Inside the vectors, there are four gene pathways. PgpdA promoter starts the transcription of HsbA, Bbchit, MCL1, Ttyptophan-MazF. TtrpC terminator terminates the transcription. It is well to be reminded that the pathway, PgpdA-Ttyptophan-MazF-TtrpC has a special switch, which induces apoptosis in the absence of tryptophan.<br /> ='''''Results'''''=<br /> ==''Adhesion:HsbA''==<br /> In this part, a strong promoter, PgpdA allows the HsbA protein to be expressed without induction. In the HsbA macro verification protocol, they could compare whether there was any change of the position and number of spores in the observing area. The wild-type Metarhizium anisopliae groups had the adherence rate in average of 26.7%. The Metarhizium anisopliae HsbA transformant groups had the adherence rate in average of 97.7%. In conclusion, this result confirmed that Metarhizium anisopliae HsbA transformant certainly enhanced the capacity of adhesion.<br /> ==''Penetration:BbChit''==<br /> In order to verify the function of Bbchit, they improved Kan Zhuo's chitin transparent circle method for verification. They stained the czapek solid medium without chitin colloids in red with 0.1% Congo red dye solution and then inoculated wild-type and transformed Metarhizium.Then they compared the size of the colony and the transparent circle between wild type and transformant. The size measured is the diameter of the chitin transparent ring (R2) and colony(R1). The ratio of the diameters showed as R2/R1.The conclusion is that the chitinase activity of transformed Metarhizium anisopliae is enhanced by 1.3 times.<br /> ==''Immune avoidance:MCL1''==<br /> In this part, PgpdA is a strong promoter that allows MCL1 to be expressed without induction. Change of nodules formed of hemocytes in cockroach hemolymph after injecting hyphae homogenate. At the time point of 0.5h, 1h and 8h, the nodules caused by M.anasopliae is significantly higher than transformants´ which means immune-avoidance occurs in genetically enhanced M.anasopliae.<br /> ==''Suicide switch''==<br /> In order to confirm the limited concentration of tryptophan, they did a macro experiment. They put Metarhizium in an L-Tryptophan concentration gradient Petrie dishes from 0.05% to 0.14% with solid and liquid czapek. It was seen that the Metarhizium can stay alive at the L-Trp concentration of 0.09% or higher while it could not grow well or die at a lower concentration. They could also see that Metarhizium could not survive without L-Tryptophan.<br /> ='''''GreenGround trapbox'''''=<br /> Using all four parts they designed a trap box named GreenGround to bring their idea into real world. They mixed spores, banana powder and oil together to form emulsifiable powder, which is applied to non-woven fabrics for use. They developed it in three generations.<br /> ==''First generation''==<br /> At first, they used oil solution to contain Metarhizium anisopliae. One area was a hollow cube to contain baits for cockroaches. There were breathable holes on top of it to make sure the smell of the baits got out. There were four entrances which allowed the cockroaches to enter. When cockroaches got in, they would get in contact with Metarhizium anisopliae which would kill them. And the cockroaches could also get out and infect other cockroaches by contacting them.<br /> After the human practice in Chongqing, they knew that cockroaches will be likely to climb gentle slope instead of entrances it.<br /> ==''Second generation''==<br /> They decided to use Metarhizium anisopliae emulsifiable powder on nonwoven fabric instead of oil solution. This time they mixed the baits with Metarhizium anisopliae emulsifiable powder and brush them on nonwoven fabric. They made a box which formed a cleft for the cockroaches to climb in through the gentle slope. In the middle were 5 magnet cubes, which could adjust the height of the cleft according to the actual size of the cockroach. They found out that magnets are not stable enough and that acrylic has its own limitations.<br /> ==''Final generation''==<br /> After they found acrylic can no longer fit their needs, they change the material into polypropylene. They also mix baits with Metarhizium anisopliae emulsifiable powder on polypropylene surface. They screw the lid down through the cylinder to form cleft which is adjustable. The cockroaches can climb through the slope and get in contact with Metarhizium anisopliae which will lead to death.<br /> ='''''Conclusion'''''=<br /> GreenGround trap box is safe and easy to use. It does not cause environmental toxicity and the development of resistance in cockroaches. Their improved M. anisopliae has better adhesion rate, is more successful in penetrating the host and can avoid immune system of the host, comparing to WT M. anisopliae.<br /> They wanted to make their product avalliable for the public since lots of poor neighborhoods in China have problem with cockroaches and other insects. They suceed in that by improving they trap box to maximize its function and to make it cheap. They also put their GreenGround box on popular app so they can get a feedback from public.<br /> ='''''Literature'''''=<br /> [1] http://2018.igem.org/Team:SZU-China<br /> <br /> [2] Mpuchane S, Matsheka IM, Gashe BA, Allotey J, Murindamombe G, Mrema N. Microbiological studies of cockroaches from three localities in Gaborone, Botswana. Afr J Food Nutr Sci. 2006;6:56–59<br /> <br /> [3] Wu X, Appel AG.J Econ Entomol. 2017 Jun 1;110(3):1203-1209<br /> <br /> [4] Wang C, Lü Dingding, Li Lin. Study on pathogenicity and degradation mechanism of entomogenous fungi [C]// Chinese Society of Fungal Sciences Academic Symposium. 2008<br /> <br /> [5] Ye Zhang, Zhongren Lei, Haihong Wang, Jiqing Zhan. Prokaryotic expression and immunolocalization Beauveria bassiana HsbA protein [J] Chinese Agricultural Sciences, 2013,46 (21): 4534-4541<br /> <br /> [6] Liu Zhihui, Chen Shouwen, Guo Zhihong, et al. Correlation between extracellular protease and chitinase activity of Beauveria bassiana and virulence to Asian corn borer[J]. Journal of Huazhong Agricultural University, 2005, 24(4) :364-368<br /> <br /> [7] Wang C, St Leger R J. A collagenous protective coat enables Metarhizium anisopliae to evade insect immune responses.[J]. Proceedings of the National Academy of Sciences of the United States of America, 2006, 103(17):6647-6652<br /> <br /> [8] Sowa S M, Keeley L L. Free amino acids in the hemolymph of the cockroach, Blaberus discoidalis[J]. Comparative Biochemistry &amp; Physiology Part A Physiology, 1996, 113(2):131</div>

Roberta Mulac https://wiki.fkkt.uni-lj.si/index.php?title=Cockroach_terminator&diff=14684 2018-12-17T17:31:58Z

<p>Roberta Mulac: </p> <hr /> <div>The SZU-China team won the golden medal for best manufacturing project and was the second runner up for the Grand prize. Web page of SZU-China team is: http://2018.igem.org/Team:SZU-China. <br /> ='''''Introduction'''''=<br /> Cockroaches are among the most common household pests, they harbor in damp and unsanitary places such as sewers, garbage disposals, kitchens, and bathroom, feed on human’s and pet’s food. Cockroaches have been implicated in the transmission of several pathogenic organisms such as E.coli and Salmonella enteritidis which can cause diarrhea, pneumonia and so on. [2]<br /> Currently, there are two ways to control the population of cockroaches, physical and chemical methods. However, a chemical method like foggers, boric acid, and gel bait may cause environmental toxicity and the development of resistance in cockroaches. A physical method such as “cockroach house”, only trap few cockroaches. Neither chemical nor physical method can’t achieve the goal of environment-friendly and efficient control. [3]<br /> ='''''Chassis and parts'''''=<br /> The SZU-China team choose a kind of entomogenous fungi, Metarhizium anisopliae, as biological chassis. It is considered a safe and prospective choice for causing disease in insects only. [4]<br /> Its infection process is divided into the following steps: <br /> <br /> 1. spore attachment <br /> <br /> 2. penetration through the body wall<br /> <br /> 3. colonization in vivo <br /> <br /> 4. killing the host<br /> <br /> 5. re-sporulation.<br /> <br /> It is a natural insecticide, and by genetically-enhancing it, we can make better use of it.<br /> But its lethality is still needed to be improved, which limits it's widespread used. They construct a system to enhance its virulence. Their system contains three parts: HsbA, BbChit, and MCL1. These three genes work sequentially during infecting cockroach, promote adhesion, penetration, and immune-avoidance respectively.<br /> HsbA- The HsbA from Beauveria bassiana (the entomopathogenic fungi, used as a biological insecticide) encodes a membrane surface hydrophobic protein A, located on the surface of fungus. This protein works like “glue” by forming a hydrophobic bond between spores and the waxy epicuticle of their host. [5]<br /> BbChit- Cockroaches’ body wall is composed of protein, chitin, and lipids. BbChit, which comes from Beauveria bassiana, encodes chitinase which can hydrolize chitin to penetrate the host. Thereby, fungus can enter into hemolymph. [6]<br /> MCL1-It encodes a collagen-like protein, which can combine with β-glucan. β-glucan is distributed on the cell wall surface of fungus and is the recognization site for insect hemocytes to combine and clear invader. MCL1 acts like putting an “invisible cloak” on the fungus, so that fungus can evade being recognized by the host immune system. Cockroaches have powerful immune system. If there is no effective response, their chassis won't be recognized and killed by host hemocytes. The MCL1 gene from Metarhizium Robertsii can encode collagen-like protein, which binds and masks β-1,3-glucan (an antigen that can be recognized by hemocytes), thus Metarhizium anisopliae can escape the immune response of cockroaches. [7]<br /> ='''''Safety mechanism'''''=<br /> They designed a suicide switch for their system. It consists of tryptophan attenuator and MazF (venom protein). Tryptophan attenuator will be switched on in high Trp concentration, while it will be switched off on the contrary. To make MazF expressed in chassis, they added a Kozak sequence in front of these two parts. They create a high Tryptophan concentration environment while culturing the fungus, and average concentration of Trp inside cockroach hemolymph is about 0.15%. In those environments, the tryptophan attenuator forms a hairpin structure that stop transcription ahead of MazF, so that fungus can stay alive. If the fungus live in the low tryptophan environment , the suicide switch will be turned on and the MazF starts to express, which leads to death of fungus. [8]<br /> ='''''Construction of plasmid vector'''''=<br /> They used the fungal plasmid pBC for their project, which can propagate in Metarhizium anisopliae. The vector contains the following parts:<br /> <br /> PgpD is the constitutive promoter of the glyceraldehyde-3-phosphate dehydrogenase gene (gpdA) from Aspergillus nidulans;<br /> TtrpC is a tryptophan terminator from Aspergillus nidulans;<br /> HsbA is encoded by the gene HsbA from Beauveria bassiana; <br /> BbChit is encoded by the gene Bbchit from Beauveria bassiana; <br /> MCL1_Metarhizium robertsii and <br /> Tryptophan-MazF-suicide switch, induces apoptosis in the absence of tryptophan.<br /> <br /> Inside the vectors, there are four gene pathways. PgpdA promoter starts the transcription of HsbA, Bbchit, MCL1, Ttyptophan-MazF. TtrpC terminator terminates the transcription. It is well to be reminded that the pathway, PgpdA-Ttyptophan-MazF-TtrpC has a special switch, which induces apoptosis in the absence of tryptophan.<br /> ='''''Results'''''=<br /> ==''Adhesion:HsbA''==<br /> In this part, a strong promoter, PgpdA allows the HsbA protein to be expressed without induction. In the HsbA macro verification protocol, they could compare whether there was any change of the position and number of spores in the observing area. The wild-type Metarhizium anisopliae groups had the adherence rate in average of 26.7%. The Metarhizium anisopliae HsbA transformant groups had the adherence rate in average of 97.7%. In conclusion, this result confirmed that Metarhizium anisopliae HsbA transformant certainly enhanced the capacity of adhesion.<br /> ==''Penetration:BbChit''==<br /> In order to verify the function of Bbchit, they improved Kan Zhuo's chitin transparent circle method for verification. They stained the czapek solid medium without chitin colloids in red with 0.1% Congo red dye solution and then inoculated wild-type and transformed Metarhizium.Then they compared the size of the colony and the transparent circle between wild type and transformant. The size measured is the diameter of the chitin transparent ring (R2) and colony(R1). The ratio of the diameters showed as R2/R1.The conclusion is that the chitinase activity of transformed Metarhizium anisopliae is enhanced by 1.3 times.<br /> ==''Immune avoidance:MCL1''==<br /> In this part, PgpdA is a strong promoter that allows MCL1 to be expressed without induction. Change of nodules formed of hemocytes in cockroach hemolymph after injecting hyphae homogenate. At the time point of 0.5h, 1h and 8h, the nodules caused by M.anasopliae is significantly higher than transformants´ which means immune-avoidance occurs in genetically enhanced M.anasopliae.<br /> ==''Suicide switch''==<br /> In order to confirm the limited concentration of tryptophan, they did a macro experiment. They put Metarhizium in an L-Tryptophan concentration gradient Petrie dishes from 0.05% to 0.14% with solid and liquid czapek. It was seen that the Metarhizium can stay alive at the L-Trp concentration of 0.09% or higher while it could not grow well or die at a lower concentration. They could also see that Metarhizium could not survive without L-Tryptophan.<br /> ='''''GreenGround trapbox'''''=<br /> Using all four parts they designed a trap box named GreenGround to bring their idea into real world. They mixed spores, banana powder and oil together to form emulsifiable powder, which is applied to non-woven fabrics for use. They developed it in three generations.<br /> ==''First generation''==<br /> At first, they used oil solution to contain Metarhizium anisopliae. One area was a hollow cube to contain baits for cockroaches. There were breathable holes on top of it to make sure the smell of the baits got out. There were four entrances which allowed the cockroaches to enter. When cockroaches got in, they would get in contact with Metarhizium anisopliae which would kill them. And the cockroaches could also get out and infect other cockroaches by contacting them.<br /> After the human practice in Chongqing, they knew that cockroaches will be likely to climb gentle slope instead of entrances it.<br /> ==''Second generation''==<br /> They decided to use Metarhizium anisopliae emulsifiable powder on nonwoven fabric instead of oil solution. This time they mixed the baits with Metarhizium anisopliae emulsifiable powder and brush them on nonwoven fabric. They made a box which formed a cleft for the cockroaches to climb in through the gentle slope. In the middle were 5 magnet cubes, which could adjust the height of the cleft according to the actual size of the cockroach. They found out that magnets are not stable enough and that acrylic has its own limitations.<br /> ==''Final generation''==<br /> After they found acrylic can no longer fit their needs, they change the material into polypropylene. They also mix baits with Metarhizium anisopliae emulsifiable powder on polypropylene surface. They screw the lid down through the cylinder to form cleft which is adjustable. The cockroaches can climb through the slope and get in contact with Metarhizium anisopliae which will lead to death.<br /> ='''''Conclusion'''''=<br /> GreenGround trap box is safe and easy to use. It does not cause environmental toxicity and the development of resistance in cockroaches. Their improved M. anisopliae has better adhesion rate, is more successful in penetrating the host and can avoid immune system of the host, comparing to WT M. anisopliae.<br /> They wanted to make their product avalliable for the public since lots of poor neighborhoods in China have problem with cockroaches and other insects. They suceed in that by improving they trap box to maximize its function and to make it cheap. They also put their GreenGround box on popular app so they can get a feedback from public.<br /> ='''''Literature'''''=<br /> [1] http://2018.igem.org/Team:SZU-China<br /> [2] Mpuchane S, Matsheka IM, Gashe BA, Allotey J, Murindamombe G, Mrema N. Microbiological studies of cockroaches from three localities in Gaborone, Botswana. Afr J Food Nutr Sci. 2006;6:56–59<br /> [3] Wu X, Appel AG.J Econ Entomol. 2017 Jun 1;110(3):1203-1209<br /> [4] Wang C, Lü Dingding, Li Lin. Study on pathogenicity and degradation mechanism of entomogenous fungi [C]// Chinese Society of Fungal Sciences Academic Symposium. 2008<br /> [5] Ye Zhang, Zhongren Lei, Haihong Wang, Jiqing Zhan. Prokaryotic expression and immunolocalization Beauveria bassiana HsbA protein [J] Chinese Agricultural Sciences, 2013,46 (21): 4534-4541<br /> [6] Liu Zhihui, Chen Shouwen, Guo Zhihong, et al. Correlation between extracellular protease and chitinase activity of Beauveria bassiana and virulence to Asian corn borer[J]. Journal of Huazhong Agricultural University, 2005, 24(4) :364-368<br /> [7] Wang C, St Leger R J. A collagenous protective coat enables Metarhizium anisopliae to evade insect immune responses.[J]. Proceedings of the National Academy of Sciences of the United States of America, 2006, 103(17):6647-6652<br /> [8] Sowa S M, Keeley L L. Free amino acids in the hemolymph of the cockroach, Blaberus discoidalis[J]. Comparative Biochemistry &amp; Physiology Part A Physiology, 1996, 113(2):131</div>

Roberta Mulac https://wiki.fkkt.uni-lj.si/index.php?title=Cockroach_terminator&diff=14682 2018-12-17T17:30:17Z

<p>Roberta Mulac: </p> <hr /> <div>The SZU-China team won the golden medal for best manufacturing project and was the second runner up for the Grand prize. Web page of SZU-China team is: http://2018.igem.org/Team:SZU-China. <br /> ='''''Introduction'''''=<br /> Cockroaches are among the most common household pests, they harbor in damp and unsanitary places such as sewers, garbage disposals, kitchens, and bathroom, feed on human’s and pet’s food. Cockroaches have been implicated in the transmission of several pathogenic organisms such as E.coli and Salmonella enteritidis which can cause diarrhea, pneumonia and so on. [2]<br /> Currently, there are two ways to control the population of cockroaches, physical and chemical methods. However, a chemical method like foggers, boric acid, and gel bait may cause environmental toxicity and the development of resistance in cockroaches. A physical method such as “cockroach house”, only trap few cockroaches. Neither chemical nor physical method can’t achieve the goal of environment-friendly and efficient control. [3]<br /> ='''''Chassis and parts'''''=<br /> The SZU-China team choose a kind of entomogenous fungi, Metarhizium anisopliae, as biological chassis. It is considered a safe and prospective choice for causing disease in insects only. [4]<br /> Its infection process is divided into the following steps: <br /> <br /> 1. spore attachment <br /> <br /> 2. penetration through the body wall<br /> <br /> 3. colonization in vivo <br /> <br /> 4. killing the host<br /> <br /> 5. re-sporulation.<br /> <br /> It is a natural insecticide, and by genetically-enhancing it, we can make better use of it.<br /> But its lethality is still needed to be improved, which limits it's widespread used. They construct a system to enhance its virulence. Their system contains three parts: HsbA, BbChit, and MCL1. These three genes work sequentially during infecting cockroach, promote adhesion, penetration, and immune-avoidance respectively.<br /> HsbA- The HsbA from Beauveria bassiana (the entomopathogenic fungi, used as a biological insecticide) encodes a membrane surface hydrophobic protein A, located on the surface of fungus. This protein works like “glue” by forming a hydrophobic bond between spores and the waxy epicuticle of their host. [5]<br /> BbChit- Cockroaches’ body wall is composed of protein, chitin, and lipids. BbChit, which comes from Beauveria bassiana, encodes chitinase which can hydrolize chitin to penetrate the host. Thereby, fungus can enter into hemolymph. [6]<br /> MCL1-It encodes a collagen-like protein, which can combine with β-glucan. β-glucan is distributed on the cell wall surface of fungus and is the recognization site for insect hemocytes to combine and clear invader. MCL1 acts like putting an “invisible cloak” on the fungus, so that fungus can evade being recognized by the host immune system. Cockroaches have powerful immune system. If there is no effective response, their chassis won't be recognized and killed by host hemocytes. The MCL1 gene from Metarhizium Robertsii can encode collagen-like protein, which binds and masks β-1,3-glucan (an antigen that can be recognized by hemocytes), thus Metarhizium anisopliae can escape the immune response of cockroaches. [7]<br /> ='''''Safety mechanism'''''=<br /> They designed a suicide switch for their system. It consists of tryptophan attenuator and MazF (venom protein). Tryptophan attenuator will be switched on in high Trp concentration, while it will be switched off on the contrary. To make MazF expressed in chassis, they added a Kozak sequence in front of these two parts. They create a high Tryptophan concentration environment while culturing the fungus, and average concentration of Trp inside cockroach hemolymph is about 0.15%. In those environments, the tryptophan attenuator forms a hairpin structure that stop transcription ahead of MazF, so that fungus can stay alive. If the fungus live in the low tryptophan environment , the suicide switch will be turned on and the MazF starts to express, which leads to death of fungus. [8]<br /> ='''''Construction of plasmid vector'''''=<br /> They used the fungal plasmid pBC for their project, which can propagate in Metarhizium anisopliae. The vector contains the following parts:<br /> <br /> PgpD is the constitutive promoter of the glyceraldehyde-3-phosphate dehydrogenase gene (gpdA) from Aspergillus nidulans;<br /> TtrpC is a tryptophan terminator from Aspergillus nidulans;<br /> HsbA is encoded by the gene HsbA from Beauveria bassiana; <br /> BbChit is encoded by the gene Bbchit from Beauveria bassiana; <br /> MCL1_Metarhizium robertsii and <br /> Tryptophan-MazF-suicide switch, induces apoptosis in the absence of tryptophan.<br /> <br /> Inside the vectors, there are four gene pathways. PgpdA promoter starts the transcription of HsbA, Bbchit, MCL1, Ttyptophan-MazF. TtrpC terminator terminates the transcription. It is well to be reminded that the pathway, PgpdA-Ttyptophan-MazF-TtrpC has a special switch, which induces apoptosis in the absence of tryptophan.<br /> ='''''Results'''''=<br /> ==''Adhesion:HsbA''==<br /> In this part, a strong promoter, PgpdA allows the HsbA protein to be expressed without induction. In the HsbA macro verification protocol, they could compare whether there was any change of the position and number of spores in the observing area. The wild-type Metarhizium anisopliae groups had the adherence rate in average of 26.7%. The Metarhizium anisopliae HsbA transformant groups had the adherence rate in average of 97.7%. In conclusion, this result confirmed that Metarhizium anisopliae HsbA transformant certainly enhanced the capacity of adhesion.<br /> ==''Penetration:BbChit''==<br /> In order to verify the function of Bbchit, they improved Kan Zhuo's chitin transparent circle method for verification. They stained the czapek solid medium without chitin colloids in red with 0.1% Congo red dye solution and then inoculated wild-type and transformed Metarhizium.Then they compared the size of the colony and the transparent circle between wild type and transformant. The size measured is the diameter of the chitin transparent ring (R2) and colony(R1). The ratio of the diameters showed as R2/R1.The conclusion is that the chitinase activity of transformed Metarhizium anisopliae is enhanced by 1.3 times.<br /> ==''Immune avoidance:MCL1''==<br /> In this part, PgpdA is a strong promoter that allows MCL1 to be expressed without induction. Change of nodules formed of hemocytes in cockroach hemolymph after injecting hyphae homogenate. At the time point of 0.5h, 1h and 8h, the nodules caused by M.anasopliae is significantly higher than transformants´ which means immune-avoidance occurs in genetically enhanced M.anasopliae.<br /> ==''Suicide switch''==<br /> In order to confirm the limited concentration of tryptophan, they did a macro experiment. They put Metarhizium in an L-Tryptophan concentration gradient Petrie dishes from 0.05% to 0.14% with solid and liquid czapek. It was seen that the Metarhizium can stay alive at the L-Trp concentration of 0.09% or higher while it could not grow well or die at a lower concentration. They could also see that Metarhizium could not survive without L-Tryptophan.<br /> ='''''GreenGround trapbox'''''=<br /> Using all four parts they designed a trap box named GreenGround to bring their idea into real world. They mixed spores, banana powder and oil together to form emulsifiable powder, which is applied to non-woven fabrics for use. They developed it in three generations.<br /> ==''First generation''==<br /> At first, they used oil solution to contain Metarhizium anisopliae. One area was a hollow cube to contain baits for cockroaches. There were breathable holes on top of it to make sure the smell of the baits got out. There were four entrances which allowed the cockroaches to enter. When cockroaches got in, they would get in contact with Metarhizium anisopliae which would kill them. And the cockroaches could also get out and infect other cockroaches by contacting them.<br /> After the human practice in Chongqing, they knew that cockroaches will be likely to climb gentle slope instead of entrances it.<br /> ==''Second generation''==<br /> They decided to use Metarhizium anisopliae emulsifiable powder on nonwoven fabric instead of oil solution. This time they mixed the baits with Metarhizium anisopliae emulsifiable powder and brush them on nonwoven fabric. They made a box which formed a cleft for the cockroaches to climb in through the gentle slope. In the middle were 5 magnet cubes, which could adjust the height of the cleft according to the actual size of the cockroach. They found out that magnets are not stable enough and that acrylic has its own limitations.<br /> ==''Final generation''==<br /> After they found acrylic can no longer fit their needs, they change the material into polypropylene. They also mix baits with Metarhizium anisopliae emulsifiable powder on polypropylene surface. They screw the lid down through the cylinder to form cleft which is adjustable. The cockroaches can climb through the slope and get in contact with Metarhizium anisopliae which will lead to death.<br /> ='''''Conclusion'''''=<br /> GreenGround trap box is safe and easy to use. It does not cause environmental toxicity and the development of resistance in cockroaches. Their improved M. anisopliae has better adhesion rate, is more successful in penetrating the host and can avoid immune system of the host, comparing to WT M. anisopliae.<br /> They wanted to make their product avalliable for the public since lots of poor neighborhoods in China have problem with cockroaches and other insects. They suceed in that by improving they trap box to maximize its function and to make it cheap. They also put their GreenGround box on popular app so they can get a feedback from public.</div>

Roberta Mulac https://wiki.fkkt.uni-lj.si/index.php?title=Cockroach_terminator&diff=14681 2018-12-17T17:29:19Z

<p>Roberta Mulac: </p> <hr /> <div>The SZU-China team won the golden medal for best manufacturing project and was the second runner up for the Grand prize. Web page of SZU-China team is: http://2018.igem.org/Team:SZU-China. <br /> ='''''Introduction'''''=<br /> Cockroaches are among the most common household pests, they harbor in damp and unsanitary places such as sewers, garbage disposals, kitchens, and bathroom, feed on human’s and pet’s food. Cockroaches have been implicated in the transmission of several pathogenic organisms such as E.coli and Salmonella enteritidis which can cause diarrhea, pneumonia and so on. [2]<br /> Currently, there are two ways to control the population of cockroaches, physical and chemical methods. However, a chemical method like foggers, boric acid, and gel bait may cause environmental toxicity and the development of resistance in cockroaches. A physical method such as “cockroach house”, only trap few cockroaches. Neither chemical nor physical method can’t achieve the goal of environment-friendly and efficient control. [3]<br /> ='''''Chassis and parts'''''=<br /> The SZU-China team choose a kind of entomogenous fungi, Metarhizium anisopliae, as biological chassis. It is considered a safe and prospective choice for causing disease in insects only. [4]<br /> Its infection process is divided into the following steps: <br /> 1. spore attachment <br /> 2. penetration through the body wall <br /> 3. colonization in vivo <br /> 4. killing the host <br /> 5. re-sporulation.<br /> It is a natural insecticide, and by genetically-enhancing it, we can make better use of it.<br /> But its lethality is still needed to be improved, which limits it's widespread used. They construct a system to enhance its virulence. Their system contains three parts: HsbA, BbChit, and MCL1. These three genes work sequentially during infecting cockroach, promote adhesion, penetration, and immune-avoidance respectively.<br /> HsbA- The HsbA from Beauveria bassiana (the entomopathogenic fungi, used as a biological insecticide) encodes a membrane surface hydrophobic protein A, located on the surface of fungus. This protein works like “glue” by forming a hydrophobic bond between spores and the waxy epicuticle of their host. [5]<br /> BbChit- Cockroaches’ body wall is composed of protein, chitin, and lipids. BbChit, which comes from Beauveria bassiana, encodes chitinase which can hydrolize chitin to penetrate the host. Thereby, fungus can enter into hemolymph. [6]<br /> MCL1-It encodes a collagen-like protein, which can combine with β-glucan. β-glucan is distributed on the cell wall surface of fungus and is the recognization site for insect hemocytes to combine and clear invader. MCL1 acts like putting an “invisible cloak” on the fungus, so that fungus can evade being recognized by the host immune system. Cockroaches have powerful immune system. If there is no effective response, their chassis won't be recognized and killed by host hemocytes. The MCL1 gene from Metarhizium Robertsii can encode collagen-like protein, which binds and masks β-1,3-glucan (an antigen that can be recognized by hemocytes), thus Metarhizium anisopliae can escape the immune response of cockroaches. [7]<br /> ='''''Safety mechanism'''''=<br /> They designed a suicide switch for their system. It consists of tryptophan attenuator and MazF (venom protein). Tryptophan attenuator will be switched on in high Trp concentration, while it will be switched off on the contrary. To make MazF expressed in chassis, they added a Kozak sequence in front of these two parts. They create a high Tryptophan concentration environment while culturing the fungus, and average concentration of Trp inside cockroach hemolymph is about 0.15%. In those environments, the tryptophan attenuator forms a hairpin structure that stop transcription ahead of MazF, so that fungus can stay alive. If the fungus live in the low tryptophan environment , the suicide switch will be turned on and the MazF starts to express, which leads to death of fungus. [8]<br /> ='''''Construction of plasmid vector'''''=<br /> They used the fungal plasmid pBC for their project, which can propagate in Metarhizium anisopliae. The vector contains the following parts:<br /> <br /> PgpD is the constitutive promoter of the glyceraldehyde-3-phosphate dehydrogenase gene (gpdA) from Aspergillus nidulans;<br /> TtrpC is a tryptophan terminator from Aspergillus nidulans;<br /> HsbA is encoded by the gene HsbA from Beauveria bassiana; <br /> BbChit is encoded by the gene Bbchit from Beauveria bassiana; <br /> MCL1_Metarhizium robertsii and <br /> Tryptophan-MazF-suicide switch, induces apoptosis in the absence of tryptophan.<br /> <br /> Inside the vectors, there are four gene pathways. PgpdA promoter starts the transcription of HsbA, Bbchit, MCL1, Ttyptophan-MazF. TtrpC terminator terminates the transcription. It is well to be reminded that the pathway, PgpdA-Ttyptophan-MazF-TtrpC has a special switch, which induces apoptosis in the absence of tryptophan.<br /> ='''''Results'''''=<br /> ==''Adhesion:HsbA''==<br /> In this part, a strong promoter, PgpdA allows the HsbA protein to be expressed without induction. In the HsbA macro verification protocol, they could compare whether there was any change of the position and number of spores in the observing area. The wild-type Metarhizium anisopliae groups had the adherence rate in average of 26.7%. The Metarhizium anisopliae HsbA transformant groups had the adherence rate in average of 97.7%. In conclusion, this result confirmed that Metarhizium anisopliae HsbA transformant certainly enhanced the capacity of adhesion.<br /> ==''Penetration:BbChit''==<br /> In order to verify the function of Bbchit, they improved Kan Zhuo's chitin transparent circle method for verification. They stained the czapek solid medium without chitin colloids in red with 0.1% Congo red dye solution and then inoculated wild-type and transformed Metarhizium.Then they compared the size of the colony and the transparent circle between wild type and transformant. The size measured is the diameter of the chitin transparent ring (R2) and colony(R1). The ratio of the diameters showed as R2/R1.The conclusion is that the chitinase activity of transformed Metarhizium anisopliae is enhanced by 1.3 times.<br /> ==''Immune avoidance:MCL1''==<br /> In this part, PgpdA is a strong promoter that allows MCL1 to be expressed without induction. Change of nodules formed of hemocytes in cockroach hemolymph after injecting hyphae homogenate. At the time point of 0.5h, 1h and 8h, the nodules caused by M.anasopliae is significantly higher than transformants´ which means immune-avoidance occurs in genetically enhanced M.anasopliae.<br /> ==''Suicide switch''==<br /> In order to confirm the limited concentration of tryptophan, they did a macro experiment. They put Metarhizium in an L-Tryptophan concentration gradient Petrie dishes from 0.05% to 0.14% with solid and liquid czapek. It was seen that the Metarhizium can stay alive at the L-Trp concentration of 0.09% or higher while it could not grow well or die at a lower concentration. They could also see that Metarhizium could not survive without L-Tryptophan.<br /> ='''''GreenGround trapbox'''''=<br /> Using all four parts they designed a trap box named GreenGround to bring their idea into real world. They mixed spores, banana powder and oil together to form emulsifiable powder, which is applied to non-woven fabrics for use. They developed it in three generations.<br /> ==''First generation''==<br /> At first, they used oil solution to contain Metarhizium anisopliae. One area was a hollow cube to contain baits for cockroaches. There were breathable holes on top of it to make sure the smell of the baits got out. There were four entrances which allowed the cockroaches to enter. When cockroaches got in, they would get in contact with Metarhizium anisopliae which would kill them. And the cockroaches could also get out and infect other cockroaches by contacting them.<br /> After the human practice in Chongqing, they knew that cockroaches will be likely to climb gentle slope instead of entrances it.<br /> ==''Second generation''==<br /> They decided to use Metarhizium anisopliae emulsifiable powder on nonwoven fabric instead of oil solution. This time they mixed the baits with Metarhizium anisopliae emulsifiable powder and brush them on nonwoven fabric. They made a box which formed a cleft for the cockroaches to climb in through the gentle slope. In the middle were 5 magnet cubes, which could adjust the height of the cleft according to the actual size of the cockroach. They found out that magnets are not stable enough and that acrylic has its own limitations.<br /> ==''Final generation''==<br /> After they found acrylic can no longer fit their needs, they change the material into polypropylene. They also mix baits with Metarhizium anisopliae emulsifiable powder on polypropylene surface. They screw the lid down through the cylinder to form cleft which is adjustable. The cockroaches can climb through the slope and get in contact with Metarhizium anisopliae which will lead to death.<br /> ='''''Conclusion'''''=<br /> GreenGround trap box is safe and easy to use. It does not cause environmental toxicity and the development of resistance in cockroaches. Their improved M. anisopliae has better adhesion rate, is more successful in penetrating the host and can avoid immune system of the host, comparing to WT M. anisopliae.<br /> They wanted to make their product avalliable for the public since lots of poor neighborhoods in China have problem with cockroaches and other insects. They suceed in that by improving they trap box to maximize its function and to make it cheap. They also put their GreenGround box on popular app so they can get a feedback from public.</div>

Roberta Mulac https://wiki.fkkt.uni-lj.si/index.php?title=Cockroach_terminator&diff=14678 2018-12-17T17:26:57Z

<p>Roberta Mulac: </p> <hr /> <div>The SZU-China team won the golden medal for best manufacturing project and was the second runner up for the Grand prize. Web page of SZU-China team is: http://2018.igem.org/Team:SZU-China. <br /> ='''''Introduction'''''=<br /> Cockroaches are among the most common household pests, they harbor in damp and unsanitary places such as sewers, garbage disposals, kitchens, and bathroom, feed on human’s and pet’s food. Cockroaches have been implicated in the transmission of several pathogenic organisms such as E.coli and Salmonella enteritidis which can cause diarrhea, pneumonia and so on. [2]<br /> Currently, there are two ways to control the population of cockroaches, physical and chemical methods. However, a chemical method like foggers, boric acid, and gel bait may cause environmental toxicity and the development of resistance in cockroaches. A physical method such as “cockroach house”, only trap few cockroaches. Neither chemical nor physical method can’t achieve the goal of environment-friendly and efficient control. [3]<br /> ='''''Chassis and parts'''''=<br /> The SZU-China team choose a kind of entomogenous fungi, Metarhizium anisopliae, as biological chassis. It is considered a safe and prospective choice for causing disease in insects only. [4]<br /> Its infection process is divided into the following steps: <br /> 1. spore attachment <br /> 2. penetration through the body wall <br /> 3. colonization in vivo <br /> 4. killing the host <br /> 5. re-sporulation.<br /> It is a natural insecticide, and by genetically-enhancing it, we can make better use of it.<br /> But its lethality is still needed to be improved, which limits it's widespread used. They construct a system to enhance its virulence. Their system contains three parts: HsbA, BbChit, and MCL1. These three genes work sequentially during infecting cockroach, promote adhesion, penetration, and immune-avoidance respectively.<br /> HsbA- The HsbA from Beauveria bassiana (the entomopathogenic fungi, used as a biological insecticide) encodes a membrane surface hydrophobic protein A, located on the surface of fungus. This protein works like “glue” by forming a hydrophobic bond between spores and the waxy epicuticle of their host. [5]<br /> BbChit- Cockroaches’ body wall is composed of protein, chitin, and lipids. BbChit, which comes from Beauveria bassiana, encodes chitinase which can hydrolize chitin to penetrate the host. Thereby, fungus can enter into hemolymph. [6]<br /> MCL1-It encodes a collagen-like protein, which can combine with β-glucan. β-glucan is distributed on the cell wall surface of fungus and is the recognization site for insect hemocytes to combine and clear invader. MCL1 acts like putting an “invisible cloak” on the fungus, so that fungus can evade being recognized by the host immune system. Cockroaches have powerful immune system. If there is no effective response, their chassis won't be recognized and killed by host hemocytes. The MCL1 gene from Metarhizium Robertsii can encode collagen-like protein, which binds and masks β-1,3-glucan (an antigen that can be recognized by hemocytes), thus Metarhizium anisopliae can escape the immune response of cockroaches. [7]<br /> ='''''Safety mechanism'''''=<br /> They designed a suicide switch for their system. It consists of tryptophan attenuator and MazF (venom protein). Tryptophan attenuator will be switched on in high Trp concentration, while it will be switched off on the contrary. To make MazF expressed in chassis, they added a Kozak sequence in front of these two parts. They create a high Tryptophan concentration environment while culturing the fungus, and average concentration of Trp inside cockroach hemolymph is about 0.15%. In those environments, the tryptophan attenuator forms a hairpin structure that stop transcription ahead of MazF, so that fungus can stay alive. If the fungus live in the low tryptophan environment , the suicide switch will be turned on and the MazF starts to express, which leads to death of fungus. [8]<br /> ='''''Construction of plasmid vector'''''=<br /> They used the fungal plasmid pBC for their project, which can propagate in Metarhizium anisopliae. The vector contains the following parts:<br /> GENE DESCRIPTION<br /> PgpdA PgpdA is the constitutive promoter of the glyceraldehyde-3-phosphate dehydrogenase gene (gpdA) from Aspergillus nidulans<br /> TtrpC TtrpC is a tryptophan terminator from Aspergillus nidulans<br /> HsbA Encoded by the gene HsbA from Beauveria bassiana <br /> BbChit Encoded by the gene Bbchit from Beauveria bassiana <br /> MCL1 MCL1_Metarhizium robertsii <br /> Tryptophan-MazF Suicide switch, induces apoptosis in the absence of tryptophan<br /> <br /> Inside the vectors, there are four gene pathways. PgpdA promoter starts the transcription of HsbA, Bbchit, MCL1, Ttyptophan-MazF. TtrpC terminator terminates the transcription. It is well to be reminded that the pathway, PgpdA-Ttyptophan-MazF-TtrpC has a special switch, which induces apoptosis in the absence of tryptophan.<br /> ='''''Results'''''=<br /> ==''Adhesion:HsbA''==<br /> In this part, a strong promoter, PgpdA allows the HsbA protein to be expressed without induction. In the HsbA macro verification protocol, they could compare whether there was any change of the position and number of spores in the observing area. The wild-type Metarhizium anisopliae groups had the adherence rate in average of 26.7%. The Metarhizium anisopliae HsbA transformant groups had the adherence rate in average of 97.7%. In conclusion, this result confirmed that Metarhizium anisopliae HsbA transformant certainly enhanced the capacity of adhesion.<br /> ==''Penetration:BbChit''==<br /> In order to verify the function of Bbchit, they improved Kan Zhuo's chitin transparent circle method for verification. They stained the czapek solid medium without chitin colloids in red with 0.1% Congo red dye solution and then inoculated wild-type and transformed Metarhizium.Then they compared the size of the colony and the transparent circle between wild type and transformant. The size measured is the diameter of the chitin transparent ring (R2) and colony(R1). The ratio of the diameters showed as R2/R1.The conclusion is that the chitinase activity of transformed Metarhizium anisopliae is enhanced by 1.3 times.<br /> ==''Immune avoidance:MCL1''==<br /> In this part, PgpdA is a strong promoter that allows MCL1 to be expressed without induction. Change of nodules formed of hemocytes in cockroach hemolymph after injecting hyphae homogenate. At the time point of 0.5h, 1h and 8h, the nodules caused by M.anasopliae is significantly higher than transformants´ which means immune-avoidance occurs in genetically enhanced M.anasopliae.<br /> ==''Suicide switch''==<br /> In order to confirm the limited concentration of tryptophan, they did a macro experiment. They put Metarhizium in an L-Tryptophan concentration gradient Petrie dishes from 0.05% to 0.14% with solid and liquid czapek. It was seen that the Metarhizium can stay alive at the L-Trp concentration of 0.09% or higher while it could not grow well or die at a lower concentration. They could also see that Metarhizium could not survive without L-Tryptophan.<br /> ='''''GreenGround trapbox'''''=<br /> Using all four parts they designed a trap box named GreenGround to bring their idea into real world. They mixed spores, banana powder and oil together to form emulsifiable powder, which is applied to non-woven fabrics for use. They developed it in three generations.<br /> ==''First generation''==<br /> At first, they used oil solution to contain Metarhizium anisopliae. One area was a hollow cube to contain baits for cockroaches. There were breathable holes on top of it to make sure the smell of the baits got out. There were four entrances which allowed the cockroaches to enter. When cockroaches got in, they would get in contact with Metarhizium anisopliae which would kill them. And the cockroaches could also get out and infect other cockroaches by contacting them.<br /> After the human practice in Chongqing, they knew that cockroaches will be likely to climb gentle slope instead of entrances it.<br /> ==''Second generation''==<br /> They decided to use Metarhizium anisopliae emulsifiable powder on nonwoven fabric instead of oil solution. This time they mixed the baits with Metarhizium anisopliae emulsifiable powder and brush them on nonwoven fabric. They made a box which formed a cleft for the cockroaches to climb in through the gentle slope. In the middle were 5 magnet cubes, which could adjust the height of the cleft according to the actual size of the cockroach. They found out that magnets are not stable enough and that acrylic has its own limitations.<br /> ==''Final generation''==<br /> After they found acrylic can no longer fit their needs, they change the material into polypropylene. They also mix baits with Metarhizium anisopliae emulsifiable powder on polypropylene surface. They screw the lid down through the cylinder to form cleft which is adjustable. The cockroaches can climb through the slope and get in contact with Metarhizium anisopliae which will lead to death.<br /> ='''''Conclusion'''''=<br /> GreenGround trap box is safe and easy to use. It does not cause environmental toxicity and the development of resistance in cockroaches. Their improved M. anisopliae has better adhesion rate, is more successful in penetrating the host and can avoid immune system of the host, comparing to WT M. anisopliae.<br /> They wanted to make their product avalliable for the public since lots of poor neighborhoods in China have problem with cockroaches and other insects. They suceed in that by improving they trap box to maximize its function and to make it cheap. They also put their GreenGround box on popular app so they can get a feedback from public.</div>

Roberta Mulac https://wiki.fkkt.uni-lj.si/index.php?title=Cockroach_terminator&diff=14674 2018-12-17T17:22:08Z

<p>Roberta Mulac: </p> <hr /> <div>The SZU-China team won the golden medal for best manufacturing project and was the second runner up for the Grand prize. Web page of SZU-China team is: http://2018.igem.org/Team:SZU-China. <br /> =Introduction=<br /> Cockroaches are among the most common household pests, they harbor in damp and unsanitary places such as sewers, garbage disposals, kitchens, and bathroom, feed on human’s and pet’s food. Cockroaches have been implicated in the transmission of several pathogenic organisms such as E.coli and Salmonella enteritidis which can cause diarrhea, pneumonia and so on. [2]<br /> Currently, there are two ways to control the population of cockroaches, physical and chemical methods. However, a chemical method like foggers, boric acid, and gel bait may cause environmental toxicity and the development of resistance in cockroaches. A physical method such as “cockroach house”, only trap few cockroaches. Neither chemical nor physical method can’t achieve the goal of environment-friendly and efficient control. [3]<br /> =Chassis and parts=<br /> The SZU-China team choose a kind of entomogenous fungi, Metarhizium anisopliae, as biological chassis. It is considered a safe and prospective choice for causing disease in insects only. [4]<br /> Its infection process is divided into the following steps: <br /> 1. spore attachment <br /> 2. penetration through the body wall <br /> 3. colonization in vivo <br /> 4. killing the host <br /> 5. re-sporulation.<br /> It is a natural insecticide, and by genetically-enhancing it, we can make better use of it.<br /> But its lethality is still needed to be improved, which limits it's widespread used. They construct a system to enhance its virulence. Their system contains three parts: HsbA, BbChit, and MCL1. These three genes work sequentially during infecting cockroach, promote adhesion, penetration, and immune-avoidance respectively.<br /> HsbA- The HsbA from Beauveria bassiana (the entomopathogenic fungi, used as a biological insecticide) encodes a membrane surface hydrophobic protein A, located on the surface of fungus. This protein works like “glue” by forming a hydrophobic bond between spores and the waxy epicuticle of their host. [5]<br /> BbChit- Cockroaches’ body wall is composed of protein, chitin, and lipids. BbChit, which comes from Beauveria bassiana, encodes chitinase which can hydrolize chitin to penetrate the host. Thereby, fungus can enter into hemolymph. [6]<br /> MCL1-It encodes a collagen-like protein, which can combine with β-glucan. β-glucan is distributed on the cell wall surface of fungus and is the recognization site for insect hemocytes to combine and clear invader. MCL1 acts like putting an “invisible cloak” on the fungus, so that fungus can evade being recognized by the host immune system. Cockroaches have powerful immune system. If there is no effective response, their chassis won't be recognized and killed by host hemocytes. The MCL1 gene from Metarhizium Robertsii can encode collagen-like protein, which binds and masks β-1,3-glucan (an antigen that can be recognized by hemocytes), thus Metarhizium anisopliae can escape the immune response of cockroaches. [7]<br /> =Safety mechanism=<br /> They designed a suicide switch for their system. It consists of tryptophan attenuator and MazF (venom protein). Tryptophan attenuator will be switched on in high Trp concentration, while it will be switched off on the contrary. To make MazF expressed in chassis, they added a Kozak sequence in front of these two parts. They create a high Tryptophan concentration environment while culturing the fungus, and average concentration of Trp inside cockroach hemolymph is about 0.15%. In those environments, the tryptophan attenuator forms a hairpin structure that stop transcription ahead of MazF, so that fungus can stay alive. If the fungus live in the low tryptophan environment , the suicide switch will be turned on and the MazF starts to express, which leads to death of fungus. [8]<br /> =Construction of plasmid vector=<br /> They used the fungal plasmid pBC for their project, which can propagate in Metarhizium anisopliae. The vector contains the following parts:<br /> GENE DESCRIPTION<br /> PgpdA PgpdA is the constitutive promoter of the glyceraldehyde-3-phosphate dehydrogenase gene (gpdA) from Aspergillus nidulans<br /> TtrpC TtrpC is a tryptophan terminator from Aspergillus nidulans<br /> HsbA Encoded by the gene HsbA from Beauveria bassiana <br /> BbChit Encoded by the gene Bbchit from Beauveria bassiana <br /> MCL1 MCL1_Metarhizium robertsii <br /> Tryptophan-MazF Suicide switch, induces apoptosis in the absence of tryptophan<br /> <br /> Inside the vectors, there are four gene pathways. PgpdA promoter starts the transcription of HsbA, Bbchit, MCL1, Ttyptophan-MazF. TtrpC terminator terminates the transcription. It is well to be reminded that the pathway, PgpdA-Ttyptophan-MazF-TtrpC has a special switch, which induces apoptosis in the absence of tryptophan.<br /> =Results=<br /> ==Adhesion:HsbA==<br /> In this part, a strong promoter, PgpdA allows the HsbA protein to be expressed without induction. In the HsbA macro verification protocol, they could compare whether there was any change of the position and number of spores in the observing area. The wild-type Metarhizium anisopliae groups had the adherence rate in average of 26.7%. The Metarhizium anisopliae HsbA transformant groups had the adherence rate in average of 97.7%. In conclusion, this result confirmed that Metarhizium anisopliae HsbA transformant certainly enhanced the capacity of adhesion.<br /> ==Penetration:BbChit==<br /> In order to verify the function of Bbchit, they improved Kan Zhuo's chitin transparent circle method for verification. They stained the czapek solid medium without chitin colloids in red with 0.1% Congo red dye solution and then inoculated wild-type and transformed Metarhizium.Then they compared the size of the colony and the transparent circle between wild type and transformant. The size measured is the diameter of the chitin transparent ring (R2) and colony(R1). The ratio of the diameters showed as R2/R1.The conclusion is that the chitinase activity of transformed Metarhizium anisopliae is enhanced by 1.3 times.<br /> ==Immune avoidance:MCL1==<br /> In this part, PgpdA is a strong promoter that allows MCL1 to be expressed without induction. Change of nodules formed of hemocytes in cockroach hemolymph after injecting hyphae homogenate. At the time point of 0.5h, 1h and 8h, the nodules caused by M.anasopliae is significantly higher than transformants´ which means immune-avoidance occurs in genetically enhanced M.anasopliae.<br /> ==Suicide switch==<br /> In order to confirm the limited concentration of tryptophan, they did a macro experiment. They put Metarhizium in an L-Tryptophan concentration gradient Petrie dishes from 0.05% to 0.14% with solid and liquid czapek. It was seen that the Metarhizium can stay alive at the L-Trp concentration of 0.09% or higher while it could not grow well or die at a lower concentration. They could also see that Metarhizium could not survive without L-Tryptophan.<br /> =GreenGround trapbox=<br /> Using all four parts they designed a trap box named GreenGround to bring their idea into real world. They mixed spores, banana powder and oil together to form emulsifiable powder, which is applied to non-woven fabrics for use. They developed it in three generations.<br /> ==First generation==<br /> At first, they used oil solution to contain Metarhizium anisopliae. One area was a hollow cube to contain baits for cockroaches. There were breathable holes on top of it to make sure the smell of the baits got out. There were four entrances which allowed the cockroaches to enter. When cockroaches got in, they would get in contact with Metarhizium anisopliae which would kill them. And the cockroaches could also get out and infect other cockroaches by contacting them.<br /> After the human practice in Chongqing, they knew that cockroaches will be likely to climb gentle slope instead of entrances it.<br /> ==Second generation==<br /> They decided to use Metarhizium anisopliae emulsifiable powder on nonwoven fabric instead of oil solution. This time they mixed the baits with Metarhizium anisopliae emulsifiable powder and brush them on nonwoven fabric. They made a box which formed a cleft for the cockroaches to climb in through the gentle slope. In the middle were 5 magnet cubes, which could adjust the height of the cleft according to the actual size of the cockroach. They found out that magnets are not stable enough and that acrylic has its own limitations.<br /> ==Final generation==<br /> After they found acrylic can no longer fit their needs, they change the material into polypropylene. They also mix baits with Metarhizium anisopliae emulsifiable powder on polypropylene surface. They screw the lid down through the cylinder to form cleft which is adjustable. The cockroaches can climb through the slope and get in contact with Metarhizium anisopliae which will lead to death.<br /> =Conclusion=<br /> GreenGround trap box is safe and easy to use. It does not cause environmental toxicity and the development of resistance in cockroaches. Their improved M. anisopliae has better adhesion rate, is more successful in penetrating the host and can avoid immune system of the host, comparing to WT M. anisopliae.<br /> They wanted to make their product avalliable for the public since lots of poor neighborhoods in China have problem with cockroaches and other insects. They suceed in that by improving they trap box to maximize its function and to make it cheap. They also put their GreenGround box on popular app so they can get a feedback from public.</div>

Roberta Mulac https://wiki.fkkt.uni-lj.si/index.php?title=Cockroach_terminator&diff=14673 2018-12-17T17:21:18Z

<p>Roberta Mulac: New page: The SZU-China team won the golden medal for best manufacturing project and was the second runner up for the Grand prize. Web page of SZU-China team is: http://2018.igem.org/Team:SZU-China....</p> <hr /> <div>The SZU-China team won the golden medal for best manufacturing project and was the second runner up for the Grand prize. Web page of SZU-China team is: http://2018.igem.org/Team:SZU-China. <br /> =Introduction=<br /> Cockroaches are among the most common household pests, they harbor in damp and unsanitary places such as sewers, garbage disposals, kitchens, and bathroom, feed on human’s and pet’s food. Cockroaches have been implicated in the transmission of several pathogenic organisms such as E.coli and Salmonella enteritidis which can cause diarrhea, pneumonia and so on. [2]<br /> Currently, there are two ways to control the population of cockroaches, physical and chemical methods. However, a chemical method like foggers, boric acid, and gel bait may cause environmental toxicity and the development of resistance in cockroaches. A physical method such as “cockroach house”, only trap few cockroaches. Neither chemical nor physical method can’t achieve the goal of environment-friendly and efficient control. [3]<br /> =Chassis and parts=<br /> The SZU-China team choose a kind of entomogenous fungi, Metarhizium anisopliae, as biological chassis. It is considered a safe and prospective choice for causing disease in insects only. [4]<br /> Its infection process is divided into the following steps: <br /> 1. spore attachment <br /> 2. penetration through the body wall <br /> 3. colonization in vivo <br /> 4. killing the host <br /> 5. re-sporulation.<br /> It is a natural insecticide, and by genetically-enhancing it, we can make better use of it.<br /> But its lethality is still needed to be improved, which limits it's widespread used. They construct a system to enhance its virulence. Their system contains three parts: HsbA, BbChit, and MCL1. These three genes work sequentially during infecting cockroach, promote adhesion, penetration, and immune-avoidance respectively.<br /> HsbA- The HsbA from Beauveria bassiana (the entomopathogenic fungi, used as a biological insecticide) encodes a membrane surface hydrophobic protein A, located on the surface of fungus. This protein works like “glue” by forming a hydrophobic bond between spores and the waxy epicuticle of their host. [5]<br /> BbChit- Cockroaches’ body wall is composed of protein, chitin, and lipids. BbChit, which comes from Beauveria bassiana, encodes chitinase which can hydrolize chitin to penetrate the host. Thereby, fungus can enter into hemolymph. [6]<br /> MCL1-It encodes a collagen-like protein, which can combine with β-glucan. β-glucan is distributed on the cell wall surface of fungus and is the recognization site for insect hemocytes to combine and clear invader. MCL1 acts like putting an “invisible cloak” on the fungus, so that fungus can evade being recognized by the host immune system. Cockroaches have powerful immune system. If there is no effective response, their chassis won't be recognized and killed by host hemocytes. The MCL1 gene from Metarhizium Robertsii can encode collagen-like protein, which binds and masks β-1,3-glucan (an antigen that can be recognized by hemocytes), thus Metarhizium anisopliae can escape the immune response of cockroaches. [7]<br /> =Safety mechanism=<br /> They designed a suicide switch for their system. It consists of tryptophan attenuator and MazF (venom protein). Tryptophan attenuator will be switched on in high Trp concentration, while it will be switched off on the contrary. To make MazF expressed in chassis, they added a Kozak sequence in front of these two parts. They create a high Tryptophan concentration environment while culturing the fungus, and average concentration of Trp inside cockroach hemolymph is about 0.15%. In those environments, the tryptophan attenuator forms a hairpin structure that stop transcription ahead of MazF, so that fungus can stay alive. If the fungus live in the low tryptophan environment , the suicide switch will be turned on and the MazF starts to express, which leads to death of fungus. [8]<br /> =Construction of plasmid vector=<br /> They used the fungal plasmid pBC for their project, which can propagate in Metarhizium anisopliae. The vector contains the following parts:<br /> GENE DESCRIPTION<br /> PgpdA PgpdA is the constitutive promoter of the glyceraldehyde-3-phosphate dehydrogenase gene (gpdA) from Aspergillus nidulans<br /> TtrpC TtrpC is a tryptophan terminator from Aspergillus nidulans<br /> HsbA Encoded by the gene HsbA from Beauveria bassiana <br /> BbChit Encoded by the gene Bbchit from Beauveria bassiana <br /> MCL1 MCL1_Metarhizium robertsii <br /> Tryptophan-MazF Suicide switch, induces apoptosis in the absence of tryptophan<br /> <br /> Inside the vectors, there are four gene pathways. PgpdA promoter starts the transcription of HsbA, Bbchit, MCL1, Ttyptophan-MazF. TtrpC terminator terminates the transcription. It is well to be reminded that the pathway, PgpdA-Ttyptophan-MazF-TtrpC has a special switch, which induces apoptosis in the absence of tryptophan.<br /> =Results=<br /> ==Adhesion:HsbA==<br /> In this part, a strong promoter, PgpdA allows the HsbA protein to be expressed without induction. In the HsbA macro verification protocol, they could compare whether there was any change of the position and number of spores in the observing area. The wild-type Metarhizium anisopliae groups had the adherence rate in average of 26.7%. The Metarhizium anisopliae HsbA transformant groups had the adherence rate in average of 97.7%. In conclusion, this result confirmed that Metarhizium anisopliae HsbA transformant certainly enhanced the capacity of adhesion.<br /> ==Penetration:BbChit==<br /> In order to verify the function of Bbchit, they improved Kan Zhuo's chitin transparent circle method for verification. They stained the czapek solid medium without chitin colloids in red with 0.1% Congo red dye solution and then inoculated wild-type and transformed Metarhizium.Then they compared the size of the colony and the transparent circle between wild type and transformant. The size measured is the diameter of the chitin transparent ring (R2) and colony(R1). The ratio of the diameters showed as R2/R1.The conclusion is that the chitinase activity of transformed Metarhizium anisopliae is enhanced by 1.3 times.<br /> ==Immune avoidance:MCL1==<br /> In this part, PgpdA is a strong promoter that allows MCL1 to be expressed without induction. Change of nodules formed of hemocytes in cockroach hemolymph after injecting hyphae homogenate. At the time point of 0.5h, 1h and 8h, the nodules caused by M.anasopliae is significantly higher than transformants´ which means immune-avoidance occurs in genetically enhanced M.anasopliae.<br /> ==Suicide switch==<br /> In order to confirm the limited concentration of tryptophan, they did a macro experiment. They put Metarhizium in an L-Tryptophan concentration gradient Petrie dishes from 0.05% to 0.14% with solid and liquid czapek. It was seen that the Metarhizium can stay alive at the L-Trp concentration of 0.09% or higher while it could not grow well or die at a lower concentration. They could also see that Metarhizium could not survive without L-Tryptophan.<br /> =GreenGround trapbox=<br /> Using all four parts they designed a trap box named GreenGround to bring their idea into real world. They mixed spores, banana powder and oil together to form emulsifiable powder, which is applied to non-woven fabrics for use. They developed it in three generations.<br /> ==First generation==<br /> At first, they used oil solution to contain Metarhizium anisopliae. One area was a hollow cube to contain baits for cockroaches. There were breathable holes on top of it to make sure the smell of the baits got out. There were four entrances which allowed the cockroaches to enter. When cockroaches got in, they would get in contact with Metarhizium anisopliae which would kill them. And the cockroaches could also get out and infect other cockroaches by contacting them.<br /> After the human practice in Chongqing, they knew that cockroaches will be likely to climb gentle slope instead of entrances it.<br /> ==Second generation=<br /> They decided to use Metarhizium anisopliae emulsifiable powder on nonwoven fabric instead of oil solution. This time they mixed the baits with Metarhizium anisopliae emulsifiable powder and brush them on nonwoven fabric. They made a box which formed a cleft for the cockroaches to climb in through the gentle slope. In the middle were 5 magnet cubes, which could adjust the height of the cleft according to the actual size of the cockroach. They found out that magnets are not stable enough and that acrylic has its own limitations.<br /> ==Final generation==<br /> After they found acrylic can no longer fit their needs, they change the material into polypropylene. They also mix baits with Metarhizium anisopliae emulsifiable powder on polypropylene surface. They screw the lid down through the cylinder to form cleft which is adjustable. The cockroaches can climb through the slope and get in contact with Metarhizium anisopliae which will lead to death.<br /> =Conclusion=<br /> GreenGround trap box is safe and easy to use. It does not cause environmental toxicity and the development of resistance in cockroaches. Their improved M. anisopliae has better adhesion rate, is more successful in penetrating the host and can avoid immune system of the host, comparing to WT M. anisopliae.<br /> They wanted to make their product avalliable for the public since lots of poor neighborhoods in China have problem with cockroaches and other insects. They suceed in that by improving they trap box to maximize its function and to make it cheap. They also put their GreenGround box on popular app so they can get a feedback from public.</div>

Roberta Mulac https://wiki.fkkt.uni-lj.si/index.php?title=Seminarji_SB_2018/19&diff=14668 2018-12-17T17:12:06Z

<p>Roberta Mulac: </p> <hr /> <div>V študijskem letu 2018/19 študentje predstavljajo naslednje teme: <br /> <br /> RAZISKOVALNI ČLANKI<br /> <br /> (Vpišite naslov seminarja v slovenščini in ga povežite z novo stranjo, kjer bo povzetek. Na tej novi strani naj bo pod naslovom povezava do izhodiščnega članka na spletu.) &lt;br&gt;<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/MoClo:_modularni_klonirni_sistem_za_standardizirano_sestavljanje_ve%C4%8Dgenskih_konstruktov MoClo: modularni klonirni sistem za standardizirano sestavljanje večgenskih konstruktov] (Valentina Levak)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/RNA-stikala_tipa_%C2%BBToehold%C2%AB:_de_novo_oblikovani_regulatorji_izra%C5%BEanja_genov RNA-stikala tipa Toehold: de novo oblikovani regulatorji izražanja genov] (Špela Malenšek)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Raznoliko_in_modelno_zasnovana_priprava_sinteti%C4%8Dnih_genskih_vezij_s_predvidenimi_lastnostmi Raznoliko in modelno zasnovana priprava sintetičnih genskih vezij s predvidenimi lastnostmi] (Matej Kolarič)<br /> <br /> [[Dispersing biofilms with engineered enzymatic bacteriophage]] (Fran Krstanović)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Nekaj_pogledov_na_sistemsko_biologijo_kvasovke Nekaj pogledov na sistemsko biologijo kvasovke] (Gašper Žun)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Organizacija_znotrajceli%C4%8Dnih_reakcij_z_razumsko_na%C4%8Drtovanimi_RNA_sestavi#Na.C4.8Drtovanje_in_sestavljanje_RNA_sestavov Organizacija znotrajceličnih reakcij z razumsko načrtovanimi RNA sestavi] (Urška Jelenovec)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Biolo%C5%A1ko_vezje_na_osnovi_RNA-interference_za_identifikacijo_specifi%C4%8Dnih_rakavih_celic Biološko vezje na osnovi RNA-interference za identifikacijo specifičnih rakavih celic] (Gašper Marinšek)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Kontrola_hitrosti_translacije_preko_pomožnega_mesta_5’-UTR:_energijski_kompromis_med_dostopnostjo%2C_selektivnim_razvijanjem_RNA-struktur_in_drsenjem_30S_ribosomske_podenote_po_RNA-strukturah Kontrola hitrosti translacije preko pomožnega mesta 5’-UTR: energijski kompromis med dostopnostjo, selektivnim razvijanjem RNA-struktur in drsenjem 30S ribosomske podenote po RNA-strukturah] (Neža Koritnik)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Preoblikovanje_genskega_skupka_za_fiksacijo_dušika_bakterije_Klebsiella_oxytoca Preoblikovanje genskega skupka za fiksacijo dušika bakterije ''Klebsiella oxytoca''] (Gašper Virant)<br /> <br /> <br /> NAGRAJENI ŠTUDENTSKI PROJEKTI <br /> <br /> (Vpišite naslov seminarja v slovenščini in ga povežite z novo stranjo, kjer bo povzetek. Na tej novi strani naj bo pod naslovom povezava do wiki strani študentske ekipe, katere projekt opisujete.) &lt;br&gt;<br /> [http://wiki.fkkt.uni-lj.si/index.php/Phactory:_proizvodnja_bakteriofagov_za_precizno_zdravljenje Phactory: proizvodnja bakteriofagov za precizno zdravljenje] (Rok Miklavčič)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Canditect:_hitra_detekcija_vaginalne_infekcije_s_Candido_albicans_z_uporabo_sistema_CRISPR/dCas9 Canditect – hitra detekcija vaginalne infekcije s Candido albicans z uporabo sistema CRISPR/dCas9] (Jerneja Ovčar)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/CAPOEIRA_-_razvoj_personaliziranega_cepiva_proti_raku_in_sistema_za_spremljanje_odziva_na_zdravljenje CAPOEIRA – razvoj personaliziranega cepiva proti raku in sistema za spremljanje odziva na zdravljenje] (Anamarija Habič)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Biotic_Blue_-_encimska_razgradnja_zdravilnih_u%C4%8Dinkovin_v_odpadnih_vodah#BIOTIC_BLUE BIOTIC BLUE - encimska razgradnja zdravilnih učinkovin v odpadnih vodah] (Tina Požun)<br /> <br /> Of CO&lt;sub&gt;2&lt;/sub&gt;urse - sistem za zmanjševanje izpustov ogljikovega dioksida v industriji (Kity Požek)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Cockroach_terminator Cockroach terminator]] (Roberta Mulac)<br /> <br /> Povzetki v slovenščini naj imajo 1200-1500 besed (viri v to vsoto ne štejejo). Predstavitev seminarja naj bo dolga 15 minut (13-17). Sledila bo razprava, ki praviloma ne bo daljša od 5 minut. <br /> <br /> ----<br /> <br /> <br /> Razpored po datumih predstavitev (pri vsakem terminu je navedeno število možnih seminarjev; vpišite ime in priimek pri dnevu, ko želite predstaviti seminar ter dopišite naslov seminarja, ki naj bo povezan s povzetkom): <br /> <br /> 22.11.&lt;br&gt; <br /> 1 &lt;br&gt;<br /> 2 Valentina Levak &lt;br&gt;<br /> <br /> 27.11.&lt;br&gt;<br /> 1 &lt;br&gt;<br /> 2 &lt;br&gt;<br /> 3 &lt;br&gt;<br /> 4 &lt;br&gt;<br /> <br /> 29.11.&lt;br&gt;<br /> 1 Matej Kolarič&lt;br&gt;<br /> 2 Špela Malenšek&lt;br&gt;<br /> 3 &lt;br&gt;<br /> 4 &lt;br&gt;<br /> <br /> 4.12.&lt;br&gt;<br /> 1 Gašper Žun&lt;br&gt;<br /> 2 Fran Krstanovic&lt;br&gt;<br /> 3 &lt;br&gt;<br /> 4 &lt;br&gt;<br /> <br /> 6.12.&lt;br&gt;<br /> 1 Urška Jelenovec&lt;br&gt;<br /> 2 Rok Miklavčič &lt;br&gt;<br /> 3 &lt;br&gt;<br /> 4 &lt;br&gt;<br /> <br /> 11.12.&lt;br&gt;<br /> 1 Jerneja Ovčar&lt;br&gt;<br /> 2 Neža Koritnik&lt;br&gt;<br /> 3 Gašper Virant&lt;br&gt;<br /> 4 Gašper Marinšek&lt;br&gt;<br /> <br /> 18.12.&lt;br&gt;<br /> 1 Tina Požun&lt;br&gt;<br /> 2 Anamarija Habič&lt;br&gt;<br /> 3 Roberta Mulac&lt;br&gt;<br /> 4 Kity Požek&lt;br&gt;<br /> <br /> 3.1.&lt;br&gt;<br /> 1 Urška Kašnik&lt;br&gt;<br /> 2 Nina Mavec&lt;br&gt;<br /> 3 Primož Tič&lt;br&gt;<br /> 4 Ernest Šprager&lt;br&gt;<br /> <br /> 8.1.&lt;br&gt;<br /> 1 Marija Atanasova&lt;br&gt;<br /> 2 Bine Tršavec&lt;br&gt;<br /> 3 Peter Pečan&lt;br&gt;<br /> 4 Tjaša Sorčan&lt;br&gt;<br /> <br /> 10.1.&lt;br&gt;<br /> 1 Špela Koren&lt;br&gt;<br /> 2 Natalija Pucihar&lt;br&gt;<br /> 3 Karmen Žbogar&lt;br&gt;<br /> 4 Uroš Zavrtanik&lt;br&gt;<br /> <br /> 15.1.&lt;br&gt;<br /> 1 Jerneja Kocutar&lt;br&gt;<br /> 2 Blaž Lebar&lt;br&gt;<br /> 3 Tadej Satler&lt;br&gt;<br /> 4 Miha Koprivnikar Krajnc&lt;br&gt;<br /> <br /> <br /> 17.1.&lt;br&gt;<br /> 1 Milena Stojkovska&lt;br&gt;<br /> 2</div>

Roberta Mulac https://wiki.fkkt.uni-lj.si/index.php?title=Seminarji_SB_2018/19&diff=14667 2018-12-17T17:11:02Z

<p>Roberta Mulac: </p> <hr /> <div>V študijskem letu 2018/19 študentje predstavljajo naslednje teme: <br /> <br /> RAZISKOVALNI ČLANKI<br /> <br /> (Vpišite naslov seminarja v slovenščini in ga povežite z novo stranjo, kjer bo povzetek. Na tej novi strani naj bo pod naslovom povezava do izhodiščnega članka na spletu.) &lt;br&gt;<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/MoClo:_modularni_klonirni_sistem_za_standardizirano_sestavljanje_ve%C4%8Dgenskih_konstruktov MoClo: modularni klonirni sistem za standardizirano sestavljanje večgenskih konstruktov] (Valentina Levak)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/RNA-stikala_tipa_%C2%BBToehold%C2%AB:_de_novo_oblikovani_regulatorji_izra%C5%BEanja_genov RNA-stikala tipa Toehold: de novo oblikovani regulatorji izražanja genov] (Špela Malenšek)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Raznoliko_in_modelno_zasnovana_priprava_sinteti%C4%8Dnih_genskih_vezij_s_predvidenimi_lastnostmi Raznoliko in modelno zasnovana priprava sintetičnih genskih vezij s predvidenimi lastnostmi] (Matej Kolarič)<br /> <br /> [[Dispersing biofilms with engineered enzymatic bacteriophage]] (Fran Krstanović)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Nekaj_pogledov_na_sistemsko_biologijo_kvasovke Nekaj pogledov na sistemsko biologijo kvasovke] (Gašper Žun)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Organizacija_znotrajceli%C4%8Dnih_reakcij_z_razumsko_na%C4%8Drtovanimi_RNA_sestavi#Na.C4.8Drtovanje_in_sestavljanje_RNA_sestavov Organizacija znotrajceličnih reakcij z razumsko načrtovanimi RNA sestavi] (Urška Jelenovec)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Biolo%C5%A1ko_vezje_na_osnovi_RNA-interference_za_identifikacijo_specifi%C4%8Dnih_rakavih_celic Biološko vezje na osnovi RNA-interference za identifikacijo specifičnih rakavih celic] (Gašper Marinšek)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Kontrola_hitrosti_translacije_preko_pomožnega_mesta_5’-UTR:_energijski_kompromis_med_dostopnostjo%2C_selektivnim_razvijanjem_RNA-struktur_in_drsenjem_30S_ribosomske_podenote_po_RNA-strukturah Kontrola hitrosti translacije preko pomožnega mesta 5’-UTR: energijski kompromis med dostopnostjo, selektivnim razvijanjem RNA-struktur in drsenjem 30S ribosomske podenote po RNA-strukturah] (Neža Koritnik)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Preoblikovanje_genskega_skupka_za_fiksacijo_dušika_bakterije_Klebsiella_oxytoca Preoblikovanje genskega skupka za fiksacijo dušika bakterije ''Klebsiella oxytoca''] (Gašper Virant)<br /> <br /> <br /> NAGRAJENI ŠTUDENTSKI PROJEKTI <br /> <br /> (Vpišite naslov seminarja v slovenščini in ga povežite z novo stranjo, kjer bo povzetek. Na tej novi strani naj bo pod naslovom povezava do wiki strani študentske ekipe, katere projekt opisujete.) &lt;br&gt;<br /> [http://wiki.fkkt.uni-lj.si/index.php/Phactory:_proizvodnja_bakteriofagov_za_precizno_zdravljenje Phactory: proizvodnja bakteriofagov za precizno zdravljenje] (Rok Miklavčič)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Canditect:_hitra_detekcija_vaginalne_infekcije_s_Candido_albicans_z_uporabo_sistema_CRISPR/dCas9 Canditect – hitra detekcija vaginalne infekcije s Candido albicans z uporabo sistema CRISPR/dCas9] (Jerneja Ovčar)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/CAPOEIRA_-_razvoj_personaliziranega_cepiva_proti_raku_in_sistema_za_spremljanje_odziva_na_zdravljenje CAPOEIRA – razvoj personaliziranega cepiva proti raku in sistema za spremljanje odziva na zdravljenje] (Anamarija Habič)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Biotic_Blue_-_encimska_razgradnja_zdravilnih_u%C4%8Dinkovin_v_odpadnih_vodah#BIOTIC_BLUE BIOTIC BLUE - encimska razgradnja zdravilnih učinkovin v odpadnih vodah] (Tina Požun)<br /> <br /> Of CO&lt;sub&gt;2&lt;/sub&gt;urse - sistem za zmanjševanje izpustov ogljikovega dioksida v industriji (Kity Požek)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Cockroach_terminator] (Roberta Mulac)<br /> <br /> Povzetki v slovenščini naj imajo 1200-1500 besed (viri v to vsoto ne štejejo). Predstavitev seminarja naj bo dolga 15 minut (13-17). Sledila bo razprava, ki praviloma ne bo daljša od 5 minut. <br /> <br /> ----<br /> <br /> <br /> Razpored po datumih predstavitev (pri vsakem terminu je navedeno število možnih seminarjev; vpišite ime in priimek pri dnevu, ko želite predstaviti seminar ter dopišite naslov seminarja, ki naj bo povezan s povzetkom): <br /> <br /> 22.11.&lt;br&gt; <br /> 1 &lt;br&gt;<br /> 2 Valentina Levak &lt;br&gt;<br /> <br /> 27.11.&lt;br&gt;<br /> 1 &lt;br&gt;<br /> 2 &lt;br&gt;<br /> 3 &lt;br&gt;<br /> 4 &lt;br&gt;<br /> <br /> 29.11.&lt;br&gt;<br /> 1 Matej Kolarič&lt;br&gt;<br /> 2 Špela Malenšek&lt;br&gt;<br /> 3 &lt;br&gt;<br /> 4 &lt;br&gt;<br /> <br /> 4.12.&lt;br&gt;<br /> 1 Gašper Žun&lt;br&gt;<br /> 2 Fran Krstanovic&lt;br&gt;<br /> 3 &lt;br&gt;<br /> 4 &lt;br&gt;<br /> <br /> 6.12.&lt;br&gt;<br /> 1 Urška Jelenovec&lt;br&gt;<br /> 2 Rok Miklavčič &lt;br&gt;<br /> 3 &lt;br&gt;<br /> 4 &lt;br&gt;<br /> <br /> 11.12.&lt;br&gt;<br /> 1 Jerneja Ovčar&lt;br&gt;<br /> 2 Neža Koritnik&lt;br&gt;<br /> 3 Gašper Virant&lt;br&gt;<br /> 4 Gašper Marinšek&lt;br&gt;<br /> <br /> 18.12.&lt;br&gt;<br /> 1 Tina Požun&lt;br&gt;<br /> 2 Anamarija Habič&lt;br&gt;<br /> 3 Roberta Mulac&lt;br&gt;<br /> 4 Kity Požek&lt;br&gt;<br /> <br /> 3.1.&lt;br&gt;<br /> 1 Urška Kašnik&lt;br&gt;<br /> 2 Nina Mavec&lt;br&gt;<br /> 3 Primož Tič&lt;br&gt;<br /> 4 Ernest Šprager&lt;br&gt;<br /> <br /> 8.1.&lt;br&gt;<br /> 1 Marija Atanasova&lt;br&gt;<br /> 2 Bine Tršavec&lt;br&gt;<br /> 3 Peter Pečan&lt;br&gt;<br /> 4 Tjaša Sorčan&lt;br&gt;<br /> <br /> 10.1.&lt;br&gt;<br /> 1 Špela Koren&lt;br&gt;<br /> 2 Natalija Pucihar&lt;br&gt;<br /> 3 Karmen Žbogar&lt;br&gt;<br /> 4 Uroš Zavrtanik&lt;br&gt;<br /> <br /> 15.1.&lt;br&gt;<br /> 1 Jerneja Kocutar&lt;br&gt;<br /> 2 Blaž Lebar&lt;br&gt;<br /> 3 Tadej Satler&lt;br&gt;<br /> 4 Miha Koprivnikar Krajnc&lt;br&gt;<br /> <br /> <br /> 17.1.&lt;br&gt;<br /> 1 Milena Stojkovska&lt;br&gt;<br /> 2</div>

Roberta Mulac https://wiki.fkkt.uni-lj.si/index.php?title=Seminarji_SB_2018/19&diff=14665 2018-12-17T17:10:31Z

<p>Roberta Mulac: </p> <hr /> <div>V študijskem letu 2018/19 študentje predstavljajo naslednje teme: <br /> <br /> RAZISKOVALNI ČLANKI<br /> <br /> (Vpišite naslov seminarja v slovenščini in ga povežite z novo stranjo, kjer bo povzetek. Na tej novi strani naj bo pod naslovom povezava do izhodiščnega članka na spletu.) &lt;br&gt;<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/MoClo:_modularni_klonirni_sistem_za_standardizirano_sestavljanje_ve%C4%8Dgenskih_konstruktov MoClo: modularni klonirni sistem za standardizirano sestavljanje večgenskih konstruktov] (Valentina Levak)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/RNA-stikala_tipa_%C2%BBToehold%C2%AB:_de_novo_oblikovani_regulatorji_izra%C5%BEanja_genov RNA-stikala tipa Toehold: de novo oblikovani regulatorji izražanja genov] (Špela Malenšek)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Raznoliko_in_modelno_zasnovana_priprava_sinteti%C4%8Dnih_genskih_vezij_s_predvidenimi_lastnostmi Raznoliko in modelno zasnovana priprava sintetičnih genskih vezij s predvidenimi lastnostmi] (Matej Kolarič)<br /> <br /> [[Dispersing biofilms with engineered enzymatic bacteriophage]] (Fran Krstanović)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Nekaj_pogledov_na_sistemsko_biologijo_kvasovke Nekaj pogledov na sistemsko biologijo kvasovke] (Gašper Žun)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Organizacija_znotrajceli%C4%8Dnih_reakcij_z_razumsko_na%C4%8Drtovanimi_RNA_sestavi#Na.C4.8Drtovanje_in_sestavljanje_RNA_sestavov Organizacija znotrajceličnih reakcij z razumsko načrtovanimi RNA sestavi] (Urška Jelenovec)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Biolo%C5%A1ko_vezje_na_osnovi_RNA-interference_za_identifikacijo_specifi%C4%8Dnih_rakavih_celic Biološko vezje na osnovi RNA-interference za identifikacijo specifičnih rakavih celic] (Gašper Marinšek)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Kontrola_hitrosti_translacije_preko_pomožnega_mesta_5’-UTR:_energijski_kompromis_med_dostopnostjo%2C_selektivnim_razvijanjem_RNA-struktur_in_drsenjem_30S_ribosomske_podenote_po_RNA-strukturah Kontrola hitrosti translacije preko pomožnega mesta 5’-UTR: energijski kompromis med dostopnostjo, selektivnim razvijanjem RNA-struktur in drsenjem 30S ribosomske podenote po RNA-strukturah] (Neža Koritnik)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Preoblikovanje_genskega_skupka_za_fiksacijo_dušika_bakterije_Klebsiella_oxytoca Preoblikovanje genskega skupka za fiksacijo dušika bakterije ''Klebsiella oxytoca''] (Gašper Virant)<br /> <br /> <br /> NAGRAJENI ŠTUDENTSKI PROJEKTI <br /> <br /> (Vpišite naslov seminarja v slovenščini in ga povežite z novo stranjo, kjer bo povzetek. Na tej novi strani naj bo pod naslovom povezava do wiki strani študentske ekipe, katere projekt opisujete.) &lt;br&gt;<br /> [http://wiki.fkkt.uni-lj.si/index.php/Phactory:_proizvodnja_bakteriofagov_za_precizno_zdravljenje Phactory: proizvodnja bakteriofagov za precizno zdravljenje] (Rok Miklavčič)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Canditect:_hitra_detekcija_vaginalne_infekcije_s_Candido_albicans_z_uporabo_sistema_CRISPR/dCas9 Canditect – hitra detekcija vaginalne infekcije s Candido albicans z uporabo sistema CRISPR/dCas9] (Jerneja Ovčar)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/CAPOEIRA_-_razvoj_personaliziranega_cepiva_proti_raku_in_sistema_za_spremljanje_odziva_na_zdravljenje CAPOEIRA – razvoj personaliziranega cepiva proti raku in sistema za spremljanje odziva na zdravljenje] (Anamarija Habič)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Biotic_Blue_-_encimska_razgradnja_zdravilnih_u%C4%8Dinkovin_v_odpadnih_vodah#BIOTIC_BLUE BIOTIC BLUE - encimska razgradnja zdravilnih učinkovin v odpadnih vodah] (Tina Požun)<br /> <br /> Of CO&lt;sub&gt;2&lt;/sub&gt;urse - sistem za zmanjševanje izpustov ogljikovega dioksida v industriji (Kity Požek)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Cockroach terminator] (Roberta Mulac)<br /> <br /> Povzetki v slovenščini naj imajo 1200-1500 besed (viri v to vsoto ne štejejo). Predstavitev seminarja naj bo dolga 15 minut (13-17). Sledila bo razprava, ki praviloma ne bo daljša od 5 minut. <br /> <br /> ----<br /> <br /> <br /> Razpored po datumih predstavitev (pri vsakem terminu je navedeno število možnih seminarjev; vpišite ime in priimek pri dnevu, ko želite predstaviti seminar ter dopišite naslov seminarja, ki naj bo povezan s povzetkom): <br /> <br /> 22.11.&lt;br&gt; <br /> 1 &lt;br&gt;<br /> 2 Valentina Levak &lt;br&gt;<br /> <br /> 27.11.&lt;br&gt;<br /> 1 &lt;br&gt;<br /> 2 &lt;br&gt;<br /> 3 &lt;br&gt;<br /> 4 &lt;br&gt;<br /> <br /> 29.11.&lt;br&gt;<br /> 1 Matej Kolarič&lt;br&gt;<br /> 2 Špela Malenšek&lt;br&gt;<br /> 3 &lt;br&gt;<br /> 4 &lt;br&gt;<br /> <br /> 4.12.&lt;br&gt;<br /> 1 Gašper Žun&lt;br&gt;<br /> 2 Fran Krstanovic&lt;br&gt;<br /> 3 &lt;br&gt;<br /> 4 &lt;br&gt;<br /> <br /> 6.12.&lt;br&gt;<br /> 1 Urška Jelenovec&lt;br&gt;<br /> 2 Rok Miklavčič &lt;br&gt;<br /> 3 &lt;br&gt;<br /> 4 &lt;br&gt;<br /> <br /> 11.12.&lt;br&gt;<br /> 1 Jerneja Ovčar&lt;br&gt;<br /> 2 Neža Koritnik&lt;br&gt;<br /> 3 Gašper Virant&lt;br&gt;<br /> 4 Gašper Marinšek&lt;br&gt;<br /> <br /> 18.12.&lt;br&gt;<br /> 1 Tina Požun&lt;br&gt;<br /> 2 Anamarija Habič&lt;br&gt;<br /> 3 Roberta Mulac&lt;br&gt;<br /> 4 Kity Požek&lt;br&gt;<br /> <br /> 3.1.&lt;br&gt;<br /> 1 Urška Kašnik&lt;br&gt;<br /> 2 Nina Mavec&lt;br&gt;<br /> 3 Primož Tič&lt;br&gt;<br /> 4 Ernest Šprager&lt;br&gt;<br /> <br /> 8.1.&lt;br&gt;<br /> 1 Marija Atanasova&lt;br&gt;<br /> 2 Bine Tršavec&lt;br&gt;<br /> 3 Peter Pečan&lt;br&gt;<br /> 4 Tjaša Sorčan&lt;br&gt;<br /> <br /> 10.1.&lt;br&gt;<br /> 1 Špela Koren&lt;br&gt;<br /> 2 Natalija Pucihar&lt;br&gt;<br /> 3 Karmen Žbogar&lt;br&gt;<br /> 4 Uroš Zavrtanik&lt;br&gt;<br /> <br /> 15.1.&lt;br&gt;<br /> 1 Jerneja Kocutar&lt;br&gt;<br /> 2 Blaž Lebar&lt;br&gt;<br /> 3 Tadej Satler&lt;br&gt;<br /> 4 Miha Koprivnikar Krajnc&lt;br&gt;<br /> <br /> <br /> 17.1.&lt;br&gt;<br /> 1 Milena Stojkovska&lt;br&gt;<br /> 2</div>

Roberta Mulac https://wiki.fkkt.uni-lj.si/index.php?title=Seminarji_SB_2018/19&diff=14664 2018-12-17T17:09:26Z

<p>Roberta Mulac: </p> <hr /> <div>V študijskem letu 2018/19 študentje predstavljajo naslednje teme: <br /> <br /> RAZISKOVALNI ČLANKI<br /> <br /> (Vpišite naslov seminarja v slovenščini in ga povežite z novo stranjo, kjer bo povzetek. Na tej novi strani naj bo pod naslovom povezava do izhodiščnega članka na spletu.) &lt;br&gt;<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/MoClo:_modularni_klonirni_sistem_za_standardizirano_sestavljanje_ve%C4%8Dgenskih_konstruktov MoClo: modularni klonirni sistem za standardizirano sestavljanje večgenskih konstruktov] (Valentina Levak)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/RNA-stikala_tipa_%C2%BBToehold%C2%AB:_de_novo_oblikovani_regulatorji_izra%C5%BEanja_genov RNA-stikala tipa Toehold: de novo oblikovani regulatorji izražanja genov] (Špela Malenšek)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Raznoliko_in_modelno_zasnovana_priprava_sinteti%C4%8Dnih_genskih_vezij_s_predvidenimi_lastnostmi Raznoliko in modelno zasnovana priprava sintetičnih genskih vezij s predvidenimi lastnostmi] (Matej Kolarič)<br /> <br /> [[Dispersing biofilms with engineered enzymatic bacteriophage]] (Fran Krstanović)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Nekaj_pogledov_na_sistemsko_biologijo_kvasovke Nekaj pogledov na sistemsko biologijo kvasovke] (Gašper Žun)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Organizacija_znotrajceli%C4%8Dnih_reakcij_z_razumsko_na%C4%8Drtovanimi_RNA_sestavi#Na.C4.8Drtovanje_in_sestavljanje_RNA_sestavov Organizacija znotrajceličnih reakcij z razumsko načrtovanimi RNA sestavi] (Urška Jelenovec)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Biolo%C5%A1ko_vezje_na_osnovi_RNA-interference_za_identifikacijo_specifi%C4%8Dnih_rakavih_celic Biološko vezje na osnovi RNA-interference za identifikacijo specifičnih rakavih celic] (Gašper Marinšek)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Kontrola_hitrosti_translacije_preko_pomožnega_mesta_5’-UTR:_energijski_kompromis_med_dostopnostjo%2C_selektivnim_razvijanjem_RNA-struktur_in_drsenjem_30S_ribosomske_podenote_po_RNA-strukturah Kontrola hitrosti translacije preko pomožnega mesta 5’-UTR: energijski kompromis med dostopnostjo, selektivnim razvijanjem RNA-struktur in drsenjem 30S ribosomske podenote po RNA-strukturah] (Neža Koritnik)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Preoblikovanje_genskega_skupka_za_fiksacijo_dušika_bakterije_Klebsiella_oxytoca Preoblikovanje genskega skupka za fiksacijo dušika bakterije ''Klebsiella oxytoca''] (Gašper Virant)<br /> <br /> <br /> NAGRAJENI ŠTUDENTSKI PROJEKTI <br /> <br /> (Vpišite naslov seminarja v slovenščini in ga povežite z novo stranjo, kjer bo povzetek. Na tej novi strani naj bo pod naslovom povezava do wiki strani študentske ekipe, katere projekt opisujete.) &lt;br&gt;<br /> [http://wiki.fkkt.uni-lj.si/index.php/Phactory:_proizvodnja_bakteriofagov_za_precizno_zdravljenje Phactory: proizvodnja bakteriofagov za precizno zdravljenje] (Rok Miklavčič)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Canditect:_hitra_detekcija_vaginalne_infekcije_s_Candido_albicans_z_uporabo_sistema_CRISPR/dCas9 Canditect – hitra detekcija vaginalne infekcije s Candido albicans z uporabo sistema CRISPR/dCas9] (Jerneja Ovčar)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/CAPOEIRA_-_razvoj_personaliziranega_cepiva_proti_raku_in_sistema_za_spremljanje_odziva_na_zdravljenje CAPOEIRA – razvoj personaliziranega cepiva proti raku in sistema za spremljanje odziva na zdravljenje] (Anamarija Habič)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Biotic_Blue_-_encimska_razgradnja_zdravilnih_u%C4%8Dinkovin_v_odpadnih_vodah#BIOTIC_BLUE BIOTIC BLUE - encimska razgradnja zdravilnih učinkovin v odpadnih vodah] (Tina Požun)<br /> <br /> Of CO&lt;sub&gt;2&lt;/sub&gt;urse - sistem za zmanjševanje izpustov ogljikovega dioksida v industriji (Kity Požek)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/ Cockroach terminator] (Roberta Mulac)<br /> <br /> Povzetki v slovenščini naj imajo 1200-1500 besed (viri v to vsoto ne štejejo). Predstavitev seminarja naj bo dolga 15 minut (13-17). Sledila bo razprava, ki praviloma ne bo daljša od 5 minut. <br /> <br /> ----<br /> <br /> <br /> Razpored po datumih predstavitev (pri vsakem terminu je navedeno število možnih seminarjev; vpišite ime in priimek pri dnevu, ko želite predstaviti seminar ter dopišite naslov seminarja, ki naj bo povezan s povzetkom): <br /> <br /> 22.11.&lt;br&gt; <br /> 1 &lt;br&gt;<br /> 2 Valentina Levak &lt;br&gt;<br /> <br /> 27.11.&lt;br&gt;<br /> 1 &lt;br&gt;<br /> 2 &lt;br&gt;<br /> 3 &lt;br&gt;<br /> 4 &lt;br&gt;<br /> <br /> 29.11.&lt;br&gt;<br /> 1 Matej Kolarič&lt;br&gt;<br /> 2 Špela Malenšek&lt;br&gt;<br /> 3 &lt;br&gt;<br /> 4 &lt;br&gt;<br /> <br /> 4.12.&lt;br&gt;<br /> 1 Gašper Žun&lt;br&gt;<br /> 2 Fran Krstanovic&lt;br&gt;<br /> 3 &lt;br&gt;<br /> 4 &lt;br&gt;<br /> <br /> 6.12.&lt;br&gt;<br /> 1 Urška Jelenovec&lt;br&gt;<br /> 2 Rok Miklavčič &lt;br&gt;<br /> 3 &lt;br&gt;<br /> 4 &lt;br&gt;<br /> <br /> 11.12.&lt;br&gt;<br /> 1 Jerneja Ovčar&lt;br&gt;<br /> 2 Neža Koritnik&lt;br&gt;<br /> 3 Gašper Virant&lt;br&gt;<br /> 4 Gašper Marinšek&lt;br&gt;<br /> <br /> 18.12.&lt;br&gt;<br /> 1 Tina Požun&lt;br&gt;<br /> 2 Anamarija Habič&lt;br&gt;<br /> 3 Roberta Mulac&lt;br&gt;<br /> 4 Kity Požek&lt;br&gt;<br /> <br /> 3.1.&lt;br&gt;<br /> 1 Urška Kašnik&lt;br&gt;<br /> 2 Nina Mavec&lt;br&gt;<br /> 3 Primož Tič&lt;br&gt;<br /> 4 Ernest Šprager&lt;br&gt;<br /> <br /> 8.1.&lt;br&gt;<br /> 1 Marija Atanasova&lt;br&gt;<br /> 2 Bine Tršavec&lt;br&gt;<br /> 3 Peter Pečan&lt;br&gt;<br /> 4 Tjaša Sorčan&lt;br&gt;<br /> <br /> 10.1.&lt;br&gt;<br /> 1 Špela Koren&lt;br&gt;<br /> 2 Natalija Pucihar&lt;br&gt;<br /> 3 Karmen Žbogar&lt;br&gt;<br /> 4 Uroš Zavrtanik&lt;br&gt;<br /> <br /> 15.1.&lt;br&gt;<br /> 1 Jerneja Kocutar&lt;br&gt;<br /> 2 Blaž Lebar&lt;br&gt;<br /> 3 Tadej Satler&lt;br&gt;<br /> 4 Miha Koprivnikar Krajnc&lt;br&gt;<br /> <br /> <br /> 17.1.&lt;br&gt;<br /> 1 Milena Stojkovska&lt;br&gt;<br /> 2</div>

Roberta Mulac https://wiki.fkkt.uni-lj.si/index.php?title=Seminarji_SB_2018/19&diff=14663 2018-12-17T17:08:50Z

<p>Roberta Mulac: </p> <hr /> <div>V študijskem letu 2018/19 študentje predstavljajo naslednje teme: <br /> <br /> RAZISKOVALNI ČLANKI<br /> <br /> (Vpišite naslov seminarja v slovenščini in ga povežite z novo stranjo, kjer bo povzetek. Na tej novi strani naj bo pod naslovom povezava do izhodiščnega članka na spletu.) &lt;br&gt;<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/MoClo:_modularni_klonirni_sistem_za_standardizirano_sestavljanje_ve%C4%8Dgenskih_konstruktov MoClo: modularni klonirni sistem za standardizirano sestavljanje večgenskih konstruktov] (Valentina Levak)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/RNA-stikala_tipa_%C2%BBToehold%C2%AB:_de_novo_oblikovani_regulatorji_izra%C5%BEanja_genov RNA-stikala tipa Toehold: de novo oblikovani regulatorji izražanja genov] (Špela Malenšek)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Raznoliko_in_modelno_zasnovana_priprava_sinteti%C4%8Dnih_genskih_vezij_s_predvidenimi_lastnostmi Raznoliko in modelno zasnovana priprava sintetičnih genskih vezij s predvidenimi lastnostmi] (Matej Kolarič)<br /> <br /> [[Dispersing biofilms with engineered enzymatic bacteriophage]] (Fran Krstanović)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Nekaj_pogledov_na_sistemsko_biologijo_kvasovke Nekaj pogledov na sistemsko biologijo kvasovke] (Gašper Žun)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Organizacija_znotrajceli%C4%8Dnih_reakcij_z_razumsko_na%C4%8Drtovanimi_RNA_sestavi#Na.C4.8Drtovanje_in_sestavljanje_RNA_sestavov Organizacija znotrajceličnih reakcij z razumsko načrtovanimi RNA sestavi] (Urška Jelenovec)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Biolo%C5%A1ko_vezje_na_osnovi_RNA-interference_za_identifikacijo_specifi%C4%8Dnih_rakavih_celic Biološko vezje na osnovi RNA-interference za identifikacijo specifičnih rakavih celic] (Gašper Marinšek)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Kontrola_hitrosti_translacije_preko_pomožnega_mesta_5’-UTR:_energijski_kompromis_med_dostopnostjo%2C_selektivnim_razvijanjem_RNA-struktur_in_drsenjem_30S_ribosomske_podenote_po_RNA-strukturah Kontrola hitrosti translacije preko pomožnega mesta 5’-UTR: energijski kompromis med dostopnostjo, selektivnim razvijanjem RNA-struktur in drsenjem 30S ribosomske podenote po RNA-strukturah] (Neža Koritnik)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Preoblikovanje_genskega_skupka_za_fiksacijo_dušika_bakterije_Klebsiella_oxytoca Preoblikovanje genskega skupka za fiksacijo dušika bakterije ''Klebsiella oxytoca''] (Gašper Virant)<br /> <br /> <br /> NAGRAJENI ŠTUDENTSKI PROJEKTI <br /> <br /> (Vpišite naslov seminarja v slovenščini in ga povežite z novo stranjo, kjer bo povzetek. Na tej novi strani naj bo pod naslovom povezava do wiki strani študentske ekipe, katere projekt opisujete.) &lt;br&gt;<br /> [http://wiki.fkkt.uni-lj.si/index.php/Phactory:_proizvodnja_bakteriofagov_za_precizno_zdravljenje Phactory: proizvodnja bakteriofagov za precizno zdravljenje] (Rok Miklavčič)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Canditect:_hitra_detekcija_vaginalne_infekcije_s_Candido_albicans_z_uporabo_sistema_CRISPR/dCas9 Canditect – hitra detekcija vaginalne infekcije s Candido albicans z uporabo sistema CRISPR/dCas9] (Jerneja Ovčar)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/CAPOEIRA_-_razvoj_personaliziranega_cepiva_proti_raku_in_sistema_za_spremljanje_odziva_na_zdravljenje CAPOEIRA – razvoj personaliziranega cepiva proti raku in sistema za spremljanje odziva na zdravljenje] (Anamarija Habič)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Biotic_Blue_-_encimska_razgradnja_zdravilnih_u%C4%8Dinkovin_v_odpadnih_vodah#BIOTIC_BLUE BIOTIC BLUE - encimska razgradnja zdravilnih učinkovin v odpadnih vodah] (Tina Požun)<br /> <br /> Of CO&lt;sub&gt;2&lt;/sub&gt;urse - sistem za zmanjševanje izpustov ogljikovega dioksida v industriji (Kity Požek)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/ Cockroaches terminator] (Roberta Mulac)<br /> <br /> Povzetki v slovenščini naj imajo 1200-1500 besed (viri v to vsoto ne štejejo). Predstavitev seminarja naj bo dolga 15 minut (13-17). Sledila bo razprava, ki praviloma ne bo daljša od 5 minut. <br /> <br /> ----<br /> <br /> <br /> Razpored po datumih predstavitev (pri vsakem terminu je navedeno število možnih seminarjev; vpišite ime in priimek pri dnevu, ko želite predstaviti seminar ter dopišite naslov seminarja, ki naj bo povezan s povzetkom): <br /> <br /> 22.11.&lt;br&gt; <br /> 1 &lt;br&gt;<br /> 2 Valentina Levak &lt;br&gt;<br /> <br /> 27.11.&lt;br&gt;<br /> 1 &lt;br&gt;<br /> 2 &lt;br&gt;<br /> 3 &lt;br&gt;<br /> 4 &lt;br&gt;<br /> <br /> 29.11.&lt;br&gt;<br /> 1 Matej Kolarič&lt;br&gt;<br /> 2 Špela Malenšek&lt;br&gt;<br /> 3 &lt;br&gt;<br /> 4 &lt;br&gt;<br /> <br /> 4.12.&lt;br&gt;<br /> 1 Gašper Žun&lt;br&gt;<br /> 2 Fran Krstanovic&lt;br&gt;<br /> 3 &lt;br&gt;<br /> 4 &lt;br&gt;<br /> <br /> 6.12.&lt;br&gt;<br /> 1 Urška Jelenovec&lt;br&gt;<br /> 2 Rok Miklavčič &lt;br&gt;<br /> 3 &lt;br&gt;<br /> 4 &lt;br&gt;<br /> <br /> 11.12.&lt;br&gt;<br /> 1 Jerneja Ovčar&lt;br&gt;<br /> 2 Neža Koritnik&lt;br&gt;<br /> 3 Gašper Virant&lt;br&gt;<br /> 4 Gašper Marinšek&lt;br&gt;<br /> <br /> 18.12.&lt;br&gt;<br /> 1 Tina Požun&lt;br&gt;<br /> 2 Anamarija Habič&lt;br&gt;<br /> 3 Roberta Mulac&lt;br&gt;<br /> 4 Kity Požek&lt;br&gt;<br /> <br /> 3.1.&lt;br&gt;<br /> 1 Urška Kašnik&lt;br&gt;<br /> 2 Nina Mavec&lt;br&gt;<br /> 3 Primož Tič&lt;br&gt;<br /> 4 Ernest Šprager&lt;br&gt;<br /> <br /> 8.1.&lt;br&gt;<br /> 1 Marija Atanasova&lt;br&gt;<br /> 2 Bine Tršavec&lt;br&gt;<br /> 3 Peter Pečan&lt;br&gt;<br /> 4 Tjaša Sorčan&lt;br&gt;<br /> <br /> 10.1.&lt;br&gt;<br /> 1 Špela Koren&lt;br&gt;<br /> 2 Natalija Pucihar&lt;br&gt;<br /> 3 Karmen Žbogar&lt;br&gt;<br /> 4 Uroš Zavrtanik&lt;br&gt;<br /> <br /> 15.1.&lt;br&gt;<br /> 1 Jerneja Kocutar&lt;br&gt;<br /> 2 Blaž Lebar&lt;br&gt;<br /> 3 Tadej Satler&lt;br&gt;<br /> 4 Miha Koprivnikar Krajnc&lt;br&gt;<br /> <br /> <br /> 17.1.&lt;br&gt;<br /> 1 Milena Stojkovska&lt;br&gt;<br /> 2</div>

Roberta Mulac https://wiki.fkkt.uni-lj.si/index.php?title=Seminarji_SB_2018/19&diff=14662 2018-12-17T17:08:10Z

<p>Roberta Mulac: </p> <hr /> <div>V študijskem letu 2018/19 študentje predstavljajo naslednje teme: <br /> <br /> RAZISKOVALNI ČLANKI<br /> <br /> (Vpišite naslov seminarja v slovenščini in ga povežite z novo stranjo, kjer bo povzetek. Na tej novi strani naj bo pod naslovom povezava do izhodiščnega članka na spletu.) &lt;br&gt;<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/MoClo:_modularni_klonirni_sistem_za_standardizirano_sestavljanje_ve%C4%8Dgenskih_konstruktov MoClo: modularni klonirni sistem za standardizirano sestavljanje večgenskih konstruktov] (Valentina Levak)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/RNA-stikala_tipa_%C2%BBToehold%C2%AB:_de_novo_oblikovani_regulatorji_izra%C5%BEanja_genov RNA-stikala tipa Toehold: de novo oblikovani regulatorji izražanja genov] (Špela Malenšek)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Raznoliko_in_modelno_zasnovana_priprava_sinteti%C4%8Dnih_genskih_vezij_s_predvidenimi_lastnostmi Raznoliko in modelno zasnovana priprava sintetičnih genskih vezij s predvidenimi lastnostmi] (Matej Kolarič)<br /> <br /> [[Dispersing biofilms with engineered enzymatic bacteriophage]] (Fran Krstanović)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Nekaj_pogledov_na_sistemsko_biologijo_kvasovke Nekaj pogledov na sistemsko biologijo kvasovke] (Gašper Žun)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Organizacija_znotrajceli%C4%8Dnih_reakcij_z_razumsko_na%C4%8Drtovanimi_RNA_sestavi#Na.C4.8Drtovanje_in_sestavljanje_RNA_sestavov Organizacija znotrajceličnih reakcij z razumsko načrtovanimi RNA sestavi] (Urška Jelenovec)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Biolo%C5%A1ko_vezje_na_osnovi_RNA-interference_za_identifikacijo_specifi%C4%8Dnih_rakavih_celic Biološko vezje na osnovi RNA-interference za identifikacijo specifičnih rakavih celic] (Gašper Marinšek)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Kontrola_hitrosti_translacije_preko_pomožnega_mesta_5’-UTR:_energijski_kompromis_med_dostopnostjo%2C_selektivnim_razvijanjem_RNA-struktur_in_drsenjem_30S_ribosomske_podenote_po_RNA-strukturah Kontrola hitrosti translacije preko pomožnega mesta 5’-UTR: energijski kompromis med dostopnostjo, selektivnim razvijanjem RNA-struktur in drsenjem 30S ribosomske podenote po RNA-strukturah] (Neža Koritnik)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Preoblikovanje_genskega_skupka_za_fiksacijo_dušika_bakterije_Klebsiella_oxytoca Preoblikovanje genskega skupka za fiksacijo dušika bakterije ''Klebsiella oxytoca''] (Gašper Virant)<br /> <br /> <br /> NAGRAJENI ŠTUDENTSKI PROJEKTI <br /> <br /> (Vpišite naslov seminarja v slovenščini in ga povežite z novo stranjo, kjer bo povzetek. Na tej novi strani naj bo pod naslovom povezava do wiki strani študentske ekipe, katere projekt opisujete.) &lt;br&gt;<br /> [http://wiki.fkkt.uni-lj.si/index.php/Phactory:_proizvodnja_bakteriofagov_za_precizno_zdravljenje Phactory: proizvodnja bakteriofagov za precizno zdravljenje] (Rok Miklavčič)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Canditect:_hitra_detekcija_vaginalne_infekcije_s_Candido_albicans_z_uporabo_sistema_CRISPR/dCas9 Canditect – hitra detekcija vaginalne infekcije s Candido albicans z uporabo sistema CRISPR/dCas9] (Jerneja Ovčar)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/CAPOEIRA_-_razvoj_personaliziranega_cepiva_proti_raku_in_sistema_za_spremljanje_odziva_na_zdravljenje CAPOEIRA – razvoj personaliziranega cepiva proti raku in sistema za spremljanje odziva na zdravljenje] (Anamarija Habič)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Biotic_Blue_-_encimska_razgradnja_zdravilnih_u%C4%8Dinkovin_v_odpadnih_vodah#BIOTIC_BLUE BIOTIC BLUE - encimska razgradnja zdravilnih učinkovin v odpadnih vodah] (Tina Požun)<br /> <br /> Of CO&lt;sub&gt;2&lt;/sub&gt;urse - sistem za zmanjševanje izpustov ogljikovega dioksida v industriji (Kity Požek)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Cockroaches terminator] (Roberta Mulac)<br /> <br /> Povzetki v slovenščini naj imajo 1200-1500 besed (viri v to vsoto ne štejejo). Predstavitev seminarja naj bo dolga 15 minut (13-17). Sledila bo razprava, ki praviloma ne bo daljša od 5 minut. <br /> <br /> ----<br /> <br /> <br /> Razpored po datumih predstavitev (pri vsakem terminu je navedeno število možnih seminarjev; vpišite ime in priimek pri dnevu, ko želite predstaviti seminar ter dopišite naslov seminarja, ki naj bo povezan s povzetkom): <br /> <br /> 22.11.&lt;br&gt; <br /> 1 &lt;br&gt;<br /> 2 Valentina Levak &lt;br&gt;<br /> <br /> 27.11.&lt;br&gt;<br /> 1 &lt;br&gt;<br /> 2 &lt;br&gt;<br /> 3 &lt;br&gt;<br /> 4 &lt;br&gt;<br /> <br /> 29.11.&lt;br&gt;<br /> 1 Matej Kolarič&lt;br&gt;<br /> 2 Špela Malenšek&lt;br&gt;<br /> 3 &lt;br&gt;<br /> 4 &lt;br&gt;<br /> <br /> 4.12.&lt;br&gt;<br /> 1 Gašper Žun&lt;br&gt;<br /> 2 Fran Krstanovic&lt;br&gt;<br /> 3 &lt;br&gt;<br /> 4 &lt;br&gt;<br /> <br /> 6.12.&lt;br&gt;<br /> 1 Urška Jelenovec&lt;br&gt;<br /> 2 Rok Miklavčič &lt;br&gt;<br /> 3 &lt;br&gt;<br /> 4 &lt;br&gt;<br /> <br /> 11.12.&lt;br&gt;<br /> 1 Jerneja Ovčar&lt;br&gt;<br /> 2 Neža Koritnik&lt;br&gt;<br /> 3 Gašper Virant&lt;br&gt;<br /> 4 Gašper Marinšek&lt;br&gt;<br /> <br /> 18.12.&lt;br&gt;<br /> 1 Tina Požun&lt;br&gt;<br /> 2 Anamarija Habič&lt;br&gt;<br /> 3 Roberta Mulac&lt;br&gt;<br /> 4 Kity Požek&lt;br&gt;<br /> <br /> 3.1.&lt;br&gt;<br /> 1 Urška Kašnik&lt;br&gt;<br /> 2 Nina Mavec&lt;br&gt;<br /> 3 Primož Tič&lt;br&gt;<br /> 4 Ernest Šprager&lt;br&gt;<br /> <br /> 8.1.&lt;br&gt;<br /> 1 Marija Atanasova&lt;br&gt;<br /> 2 Bine Tršavec&lt;br&gt;<br /> 3 Peter Pečan&lt;br&gt;<br /> 4 Tjaša Sorčan&lt;br&gt;<br /> <br /> 10.1.&lt;br&gt;<br /> 1 Špela Koren&lt;br&gt;<br /> 2 Natalija Pucihar&lt;br&gt;<br /> 3 Karmen Žbogar&lt;br&gt;<br /> 4 Uroš Zavrtanik&lt;br&gt;<br /> <br /> 15.1.&lt;br&gt;<br /> 1 Jerneja Kocutar&lt;br&gt;<br /> 2 Blaž Lebar&lt;br&gt;<br /> 3 Tadej Satler&lt;br&gt;<br /> 4 Miha Koprivnikar Krajnc&lt;br&gt;<br /> <br /> <br /> 17.1.&lt;br&gt;<br /> 1 Milena Stojkovska&lt;br&gt;<br /> 2</div>

Roberta Mulac https://wiki.fkkt.uni-lj.si/index.php?title=Seminarji_SB_2018/19&diff=14660 2018-12-17T17:07:23Z

<p>Roberta Mulac: </p> <hr /> <div>V študijskem letu 2018/19 študentje predstavljajo naslednje teme: <br /> <br /> RAZISKOVALNI ČLANKI<br /> <br /> (Vpišite naslov seminarja v slovenščini in ga povežite z novo stranjo, kjer bo povzetek. Na tej novi strani naj bo pod naslovom povezava do izhodiščnega članka na spletu.) &lt;br&gt;<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/MoClo:_modularni_klonirni_sistem_za_standardizirano_sestavljanje_ve%C4%8Dgenskih_konstruktov MoClo: modularni klonirni sistem za standardizirano sestavljanje večgenskih konstruktov] (Valentina Levak)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/RNA-stikala_tipa_%C2%BBToehold%C2%AB:_de_novo_oblikovani_regulatorji_izra%C5%BEanja_genov RNA-stikala tipa Toehold: de novo oblikovani regulatorji izražanja genov] (Špela Malenšek)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Raznoliko_in_modelno_zasnovana_priprava_sinteti%C4%8Dnih_genskih_vezij_s_predvidenimi_lastnostmi Raznoliko in modelno zasnovana priprava sintetičnih genskih vezij s predvidenimi lastnostmi] (Matej Kolarič)<br /> <br /> [[Dispersing biofilms with engineered enzymatic bacteriophage]] (Fran Krstanović)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Nekaj_pogledov_na_sistemsko_biologijo_kvasovke Nekaj pogledov na sistemsko biologijo kvasovke] (Gašper Žun)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Organizacija_znotrajceli%C4%8Dnih_reakcij_z_razumsko_na%C4%8Drtovanimi_RNA_sestavi#Na.C4.8Drtovanje_in_sestavljanje_RNA_sestavov Organizacija znotrajceličnih reakcij z razumsko načrtovanimi RNA sestavi] (Urška Jelenovec)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Biolo%C5%A1ko_vezje_na_osnovi_RNA-interference_za_identifikacijo_specifi%C4%8Dnih_rakavih_celic Biološko vezje na osnovi RNA-interference za identifikacijo specifičnih rakavih celic] (Gašper Marinšek)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Kontrola_hitrosti_translacije_preko_pomožnega_mesta_5’-UTR:_energijski_kompromis_med_dostopnostjo%2C_selektivnim_razvijanjem_RNA-struktur_in_drsenjem_30S_ribosomske_podenote_po_RNA-strukturah Kontrola hitrosti translacije preko pomožnega mesta 5’-UTR: energijski kompromis med dostopnostjo, selektivnim razvijanjem RNA-struktur in drsenjem 30S ribosomske podenote po RNA-strukturah] (Neža Koritnik)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Preoblikovanje_genskega_skupka_za_fiksacijo_dušika_bakterije_Klebsiella_oxytoca Preoblikovanje genskega skupka za fiksacijo dušika bakterije ''Klebsiella oxytoca''] (Gašper Virant)<br /> <br /> <br /> NAGRAJENI ŠTUDENTSKI PROJEKTI <br /> <br /> (Vpišite naslov seminarja v slovenščini in ga povežite z novo stranjo, kjer bo povzetek. Na tej novi strani naj bo pod naslovom povezava do wiki strani študentske ekipe, katere projekt opisujete.) &lt;br&gt;<br /> [http://wiki.fkkt.uni-lj.si/index.php/Phactory:_proizvodnja_bakteriofagov_za_precizno_zdravljenje Phactory: proizvodnja bakteriofagov za precizno zdravljenje] (Rok Miklavčič)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Canditect:_hitra_detekcija_vaginalne_infekcije_s_Candido_albicans_z_uporabo_sistema_CRISPR/dCas9 Canditect – hitra detekcija vaginalne infekcije s Candido albicans z uporabo sistema CRISPR/dCas9] (Jerneja Ovčar)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/CAPOEIRA_-_razvoj_personaliziranega_cepiva_proti_raku_in_sistema_za_spremljanje_odziva_na_zdravljenje CAPOEIRA – razvoj personaliziranega cepiva proti raku in sistema za spremljanje odziva na zdravljenje] (Anamarija Habič)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Biotic_Blue_-_encimska_razgradnja_zdravilnih_u%C4%8Dinkovin_v_odpadnih_vodah#BIOTIC_BLUE BIOTIC BLUE - encimska razgradnja zdravilnih učinkovin v odpadnih vodah] (Tina Požun)<br /> <br /> Of CO&lt;sub&gt;2&lt;/sub&gt;urse - sistem za zmanjševanje izpustov ogljikovega dioksida v industriji (Kity Požek)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/2018.igem.org/Team:SZU-China Cockroaches terminator] (Roberta Mulac)<br /> <br /> Povzetki v slovenščini naj imajo 1200-1500 besed (viri v to vsoto ne štejejo). Predstavitev seminarja naj bo dolga 15 minut (13-17). Sledila bo razprava, ki praviloma ne bo daljša od 5 minut. <br /> <br /> ----<br /> <br /> <br /> Razpored po datumih predstavitev (pri vsakem terminu je navedeno število možnih seminarjev; vpišite ime in priimek pri dnevu, ko želite predstaviti seminar ter dopišite naslov seminarja, ki naj bo povezan s povzetkom): <br /> <br /> 22.11.&lt;br&gt; <br /> 1 &lt;br&gt;<br /> 2 Valentina Levak &lt;br&gt;<br /> <br /> 27.11.&lt;br&gt;<br /> 1 &lt;br&gt;<br /> 2 &lt;br&gt;<br /> 3 &lt;br&gt;<br /> 4 &lt;br&gt;<br /> <br /> 29.11.&lt;br&gt;<br /> 1 Matej Kolarič&lt;br&gt;<br /> 2 Špela Malenšek&lt;br&gt;<br /> 3 &lt;br&gt;<br /> 4 &lt;br&gt;<br /> <br /> 4.12.&lt;br&gt;<br /> 1 Gašper Žun&lt;br&gt;<br /> 2 Fran Krstanovic&lt;br&gt;<br /> 3 &lt;br&gt;<br /> 4 &lt;br&gt;<br /> <br /> 6.12.&lt;br&gt;<br /> 1 Urška Jelenovec&lt;br&gt;<br /> 2 Rok Miklavčič &lt;br&gt;<br /> 3 &lt;br&gt;<br /> 4 &lt;br&gt;<br /> <br /> 11.12.&lt;br&gt;<br /> 1 Jerneja Ovčar&lt;br&gt;<br /> 2 Neža Koritnik&lt;br&gt;<br /> 3 Gašper Virant&lt;br&gt;<br /> 4 Gašper Marinšek&lt;br&gt;<br /> <br /> 18.12.&lt;br&gt;<br /> 1 Tina Požun&lt;br&gt;<br /> 2 Anamarija Habič&lt;br&gt;<br /> 3 Roberta Mulac&lt;br&gt;<br /> 4 Kity Požek&lt;br&gt;<br /> <br /> 3.1.&lt;br&gt;<br /> 1 Urška Kašnik&lt;br&gt;<br /> 2 Nina Mavec&lt;br&gt;<br /> 3 Primož Tič&lt;br&gt;<br /> 4 Ernest Šprager&lt;br&gt;<br /> <br /> 8.1.&lt;br&gt;<br /> 1 Marija Atanasova&lt;br&gt;<br /> 2 Bine Tršavec&lt;br&gt;<br /> 3 Peter Pečan&lt;br&gt;<br /> 4 Tjaša Sorčan&lt;br&gt;<br /> <br /> 10.1.&lt;br&gt;<br /> 1 Špela Koren&lt;br&gt;<br /> 2 Natalija Pucihar&lt;br&gt;<br /> 3 Karmen Žbogar&lt;br&gt;<br /> 4 Uroš Zavrtanik&lt;br&gt;<br /> <br /> 15.1.&lt;br&gt;<br /> 1 Jerneja Kocutar&lt;br&gt;<br /> 2 Blaž Lebar&lt;br&gt;<br /> 3 Tadej Satler&lt;br&gt;<br /> 4 Miha Koprivnikar Krajnc&lt;br&gt;<br /> <br /> <br /> 17.1.&lt;br&gt;<br /> 1 Milena Stojkovska&lt;br&gt;<br /> 2</div>

Roberta Mulac https://wiki.fkkt.uni-lj.si/index.php?title=Seminarji_SB_2018/19&diff=14659 2018-12-17T17:05:02Z

<p>Roberta Mulac: </p> <hr /> <div>V študijskem letu 2018/19 študentje predstavljajo naslednje teme: <br /> <br /> RAZISKOVALNI ČLANKI<br /> <br /> (Vpišite naslov seminarja v slovenščini in ga povežite z novo stranjo, kjer bo povzetek. Na tej novi strani naj bo pod naslovom povezava do izhodiščnega članka na spletu.) &lt;br&gt;<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/MoClo:_modularni_klonirni_sistem_za_standardizirano_sestavljanje_ve%C4%8Dgenskih_konstruktov MoClo: modularni klonirni sistem za standardizirano sestavljanje večgenskih konstruktov] (Valentina Levak)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/RNA-stikala_tipa_%C2%BBToehold%C2%AB:_de_novo_oblikovani_regulatorji_izra%C5%BEanja_genov RNA-stikala tipa Toehold: de novo oblikovani regulatorji izražanja genov] (Špela Malenšek)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Raznoliko_in_modelno_zasnovana_priprava_sinteti%C4%8Dnih_genskih_vezij_s_predvidenimi_lastnostmi Raznoliko in modelno zasnovana priprava sintetičnih genskih vezij s predvidenimi lastnostmi] (Matej Kolarič)<br /> <br /> [[Dispersing biofilms with engineered enzymatic bacteriophage]] (Fran Krstanović)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Nekaj_pogledov_na_sistemsko_biologijo_kvasovke Nekaj pogledov na sistemsko biologijo kvasovke] (Gašper Žun)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Organizacija_znotrajceli%C4%8Dnih_reakcij_z_razumsko_na%C4%8Drtovanimi_RNA_sestavi#Na.C4.8Drtovanje_in_sestavljanje_RNA_sestavov Organizacija znotrajceličnih reakcij z razumsko načrtovanimi RNA sestavi] (Urška Jelenovec)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Biolo%C5%A1ko_vezje_na_osnovi_RNA-interference_za_identifikacijo_specifi%C4%8Dnih_rakavih_celic Biološko vezje na osnovi RNA-interference za identifikacijo specifičnih rakavih celic] (Gašper Marinšek)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Kontrola_hitrosti_translacije_preko_pomožnega_mesta_5’-UTR:_energijski_kompromis_med_dostopnostjo%2C_selektivnim_razvijanjem_RNA-struktur_in_drsenjem_30S_ribosomske_podenote_po_RNA-strukturah Kontrola hitrosti translacije preko pomožnega mesta 5’-UTR: energijski kompromis med dostopnostjo, selektivnim razvijanjem RNA-struktur in drsenjem 30S ribosomske podenote po RNA-strukturah] (Neža Koritnik)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Preoblikovanje_genskega_skupka_za_fiksacijo_dušika_bakterije_Klebsiella_oxytoca Preoblikovanje genskega skupka za fiksacijo dušika bakterije ''Klebsiella oxytoca''] (Gašper Virant)<br /> <br /> <br /> NAGRAJENI ŠTUDENTSKI PROJEKTI <br /> <br /> (Vpišite naslov seminarja v slovenščini in ga povežite z novo stranjo, kjer bo povzetek. Na tej novi strani naj bo pod naslovom povezava do wiki strani študentske ekipe, katere projekt opisujete.) &lt;br&gt;<br /> [http://wiki.fkkt.uni-lj.si/index.php/Phactory:_proizvodnja_bakteriofagov_za_precizno_zdravljenje Phactory: proizvodnja bakteriofagov za precizno zdravljenje] (Rok Miklavčič)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Canditect:_hitra_detekcija_vaginalne_infekcije_s_Candido_albicans_z_uporabo_sistema_CRISPR/dCas9 Canditect – hitra detekcija vaginalne infekcije s Candido albicans z uporabo sistema CRISPR/dCas9] (Jerneja Ovčar)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/CAPOEIRA_-_razvoj_personaliziranega_cepiva_proti_raku_in_sistema_za_spremljanje_odziva_na_zdravljenje CAPOEIRA – razvoj personaliziranega cepiva proti raku in sistema za spremljanje odziva na zdravljenje] (Anamarija Habič)<br /> <br /> [http://wiki.fkkt.uni-lj.si/index.php/Biotic_Blue_-_encimska_razgradnja_zdravilnih_u%C4%8Dinkovin_v_odpadnih_vodah#BIOTIC_BLUE BIOTIC BLUE - encimska razgradnja zdravilnih učinkovin v odpadnih vodah] (Tina Požun)<br /> <br /> Of CO&lt;sub&gt;2&lt;/sub&gt;urse - sistem za zmanjševanje izpustov ogljikovega dioksida v industriji (Kity Požek)<br /> <br /> [http://2018.igem.org/Team:SZU-China] Cockroaches terminator<br /> <br /> Povzetki v slovenščini naj imajo 1200-1500 besed (viri v to vsoto ne štejejo). Predstavitev seminarja naj bo dolga 15 minut (13-17). Sledila bo razprava, ki praviloma ne bo daljša od 5 minut. <br /> <br /> ----<br /> <br /> <br /> Razpored po datumih predstavitev (pri vsakem terminu je navedeno število možnih seminarjev; vpišite ime in priimek pri dnevu, ko želite predstaviti seminar ter dopišite naslov seminarja, ki naj bo povezan s povzetkom): <br /> <br /> 22.11.&lt;br&gt; <br /> 1 &lt;br&gt;<br /> 2 Valentina Levak &lt;br&gt;<br /> <br /> 27.11.&lt;br&gt;<br /> 1 &lt;br&gt;<br /> 2 &lt;br&gt;<br /> 3 &lt;br&gt;<br /> 4 &lt;br&gt;<br /> <br /> 29.11.&lt;br&gt;<br /> 1 Matej Kolarič&lt;br&gt;<br /> 2 Špela Malenšek&lt;br&gt;<br /> 3 &lt;br&gt;<br /> 4 &lt;br&gt;<br /> <br /> 4.12.&lt;br&gt;<br /> 1 Gašper Žun&lt;br&gt;<br /> 2 Fran Krstanovic&lt;br&gt;<br /> 3 &lt;br&gt;<br /> 4 &lt;br&gt;<br /> <br /> 6.12.&lt;br&gt;<br /> 1 Urška Jelenovec&lt;br&gt;<br /> 2 Rok Miklavčič &lt;br&gt;<br /> 3 &lt;br&gt;<br /> 4 &lt;br&gt;<br /> <br /> 11.12.&lt;br&gt;<br /> 1 Jerneja Ovčar&lt;br&gt;<br /> 2 Neža Koritnik&lt;br&gt;<br /> 3 Gašper Virant&lt;br&gt;<br /> 4 Gašper Marinšek&lt;br&gt;<br /> <br /> 18.12.&lt;br&gt;<br /> 1 Tina Požun&lt;br&gt;<br /> 2 Anamarija Habič&lt;br&gt;<br /> 3 Roberta Mulac&lt;br&gt;<br /> 4 Kity Požek&lt;br&gt;<br /> <br /> 3.1.&lt;br&gt;<br /> 1 Urška Kašnik&lt;br&gt;<br /> 2 Nina Mavec&lt;br&gt;<br /> 3 Primož Tič&lt;br&gt;<br /> 4 Ernest Šprager&lt;br&gt;<br /> <br /> 8.1.&lt;br&gt;<br /> 1 Marija Atanasova&lt;br&gt;<br /> 2 Bine Tršavec&lt;br&gt;<br /> 3 Peter Pečan&lt;br&gt;<br /> 4 Tjaša Sorčan&lt;br&gt;<br /> <br /> 10.1.&lt;br&gt;<br /> 1 Špela Koren&lt;br&gt;<br /> 2 Natalija Pucihar&lt;br&gt;<br /> 3 Karmen Žbogar&lt;br&gt;<br /> 4 Uroš Zavrtanik&lt;br&gt;<br /> <br /> 15.1.&lt;br&gt;<br /> 1 Jerneja Kocutar&lt;br&gt;<br /> 2 Blaž Lebar&lt;br&gt;<br /> 3 Tadej Satler&lt;br&gt;<br /> 4 Miha Koprivnikar Krajnc&lt;br&gt;<br /> <br /> <br /> 17.1.&lt;br&gt;<br /> 1 Milena Stojkovska&lt;br&gt;<br /> 2</div>

Roberta Mulac https://wiki.fkkt.uni-lj.si/index.php?title=Dispersing_biofilms_with_engineered_enzymatic_bacteriophage&diff=14655 2018-12-17T16:58:30Z

<p>Roberta Mulac: /* Introduction */</p> <hr /> <div>Izvorni članek: Lu, T., et al, Dispersing biofilms with engineered enzymatic bacteriophage, PNAS, 2007.&lt;ref&gt;[http://www.pnas.org/content/104/27/11197]&lt;/ref&gt;<br /> <br /> == Introduction ==<br /> <br /> When faced with certain challenges in various living habitats, bacteria have the ability to form biofilms, or organized aggregates of microorganisms living within an extracellular polymeric matrix (EPM) &lt;ref name=&quot;jamal&quot;&gt;https://www.ncbi.nlm.nih.gov/pubmed/29042186&lt;/ref&gt;. The complex EPM is formed by heterogeneous extracellular polymeric substances (EPS), which is occupied mostly by water (97%) and other macromolecules in lower concentrations (proteins (~2%), polysaccharides (1-2%); nucleic acids (&lt;1%) and ions (bound and free)) &lt;ref name=&quot;jamal&quot; /&gt;. This organized communities are formed on either biological or non-biological surfaces, and allow bacterium to surpass harsh environmental conditions, such as UV exposure, metal toxicity, acid exposure, dehydration and salinity, phagocytosis and several antibiotics and antimicrobial agents &lt;ref name=&quot;dva&quot;&gt;https://www.ncbi.nlm.nih.gov/pubmed/15040259&lt;/ref&gt;. The ability to surpass latter conditions represent a challenge in the medical, industrial and food branches, as biofilm formation accounts for over 65% of microbial infections, and over 80% of chronic infections based on the statistics from the National Institutes of Health &lt;ref name=&quot;dva&quot; /&gt;. Due to the concerning numbers, and the rise of antibiotic resistance, a novel and effective treatment for bacterial biofilms is necessary. The main target in biofilm degradation is disruption of the EPM, more precisely to target the EPS and mechanism involved in EPS production and secretion (DNase, exopolysaccharides, protein components, cGMP/cAMP levels, signal and secretion pathways) &lt;ref&gt;[https://www.ncbi.nlm.nih.gov/pubmed/28944770]&lt;/ref&gt;. In the following paper, we will show a potential treatment with synthetically engineered bacteriophages that possess the enzymatic ability to disperse bacterial biofilms.<br /> <br /> == Bacteriophages and biofilms ==<br /> <br /> The use of bacteriophages against bacterial infections is not a novelty method, as it is dates from the early 20th century. With the growing knowledge of engineering and manipulating biological organisms, and the highly annotated phage genome, makes bacteriophages prime candidates for targeting biofilms. Bacteriophages are viruses that infect and replicate within bacteria, and in comparison with antibiotics and other antimicrobial agents, possess the ability to penetrate biofilms, and disperse biofilms by various proposed mechanisms &lt;ref&gt;[https://www.ncbi.nlm.nih.gov/pubmed/23306440]&lt;/ref&gt; &lt;ref&gt;[https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4790368/]&lt;/ref&gt;. As mentioned above, enzymatic targeting exopolysaccharides with EPS-degrading is one of the possible strategies for targeting biofilms &lt;ref name=&quot;tri&quot; /&gt;. The challenge lies in isolating a natural phage that is both specific for the bacteria to be targeted and expresses a relevant EPS-degrading enzyme. The solution lies in designing an artificial biofilm-degrading bacteriophage that express a specific EPS-degrading enzyme. <br /> <br /> == Dispersin B==<br /> <br /> While studying the strain A. actinomycetemcomitans which causes periodontal disease in adolescents and its biofilm formation and degradation properties for disease spreading, researchers have come upon a novel specific biofilm-releasing glycoside hydrolase &lt;ref name=&quot;tri&quot;&gt;https://www.ncbi.nlm.nih.gov/pubmed/15878175&lt;/ref&gt;. This novel protein disperin B or DspB has the ability to degrade an important EPS polysaccharide adhesin known as β-1,6-N-acetyl-D-glucosamine &lt;ref name=&quot;tri&quot; /&gt;. N-acetyl-D-glucosamine residues form various polymeric structures with their linear β-1,6-linkages, such as PIA, PNAG, PGA (abbreviations)&lt;ref name=&quot;tri&quot; /&gt;. By hydrolyzing polymers, the protein disrupts the formation of the biofilm matrix and allows adherent cells to be released &lt;ref name=&quot;tri&quot; /&gt;. From a structural point of view, disperin B consist of a single domain with intertwining α/β structures &lt;ref name=&quot;tri&quot; /&gt;. The protein is a member of the 20 β-hexosaminidases family (GH-20), and has a highly conserved acidic active site (D183, E184, E332) which cleaves terminal monosaccharide residues from the non-reducing end of the polymers &lt;ref name=&quot;tri&quot; /&gt;.<br /> <br /> == Design of enzymatically active bacteriophage ==<br /> <br /> Once a suitable protein for biofilm removal that covers a wide specter was found, the design of engineered bacteriophages could commence. The idea is based on exploiting the lytic phage life cycle, which is based on hijacking the cell machinery, and synthesizing components of the phage genome &lt;ref name=&quot;cet&quot;&gt;https://www.ncbi.nlm.nih.gov/pubmed/12776216&lt;/ref&gt;. Once all sufficient components are available, the phages reassembles inside the cell, causing the cell to burst and releasing its component in to the local environment &lt;ref name=&quot;cet&quot; /&gt;. This two-pronged attack strategy, would exploit the enzyme to remove bacterial biofilm, and the phage infections to lyse cells, while achieving high concentrations of the enzyme and lytic phage. The backbone of the design is based on using an E.coli specific lytic T7 phage, which was modified in a way that it had a few of nonessential gene deletions. The T7 phage had one of the first completely sequenced genomes (40-kb) that codes for 55 proteins &lt;ref name=&quot;pet&quot;&gt;https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2525648/&lt;/ref&gt;. The T7 is widely used in molecular biology and has various traits that make this strain suitable for phage experiments &lt;ref name=&quot;pet&quot; /&gt;. The biofilm removing T7 phage was design to express DspB under the strong control of T7 ϕ10 promoter intracellularly during the infection, so that it could be released in to the environment upon cell lysis. The experiment was focused on strains that possess the F-plasmid, as it enhances biofilm maturation, and forms more thick biofilms, making them a more appealing group. Bacterial strains that contain the F-plasmid can disrupt efficient T7 replication. To tackle this problem, they inserted a 1.2 gene from T3 phage so the phage would not be limited only to strains that lack the F-plasmid, but to widen the spectrum of target &lt;ref name=&quot;set&quot;&gt;https://www.ncbi.nlm.nih.gov/pmc/articles/PMC208987/&lt;/ref&gt;. Gene 1.2 is an inhibitor of the host dGTPase, which is involved in the process of DNA replication &lt;ref name=&quot;set&quot; /&gt;. A control was designed by cloning an S-tag into the T7 genome, to assure quality results. The phages were named T7DspB and T7Control respectively.<br /> <br /> == Characterization of enzymatically active bacteriophage ==<br /> <br /> The first task was to determine if T7DspB was more effective against biofilms than T7Control. Effectiveness of modified phages was also compared to wild-type T3 and T7 phages to determine if modified phages possess an edge over natural occurring ones. To measure effectiveness, crystal violet (CV) methods was used, which is based on staining attached cells (in our case bacterial cells/biofilms) with crystal violet dye, which binds to proteins and DNA &lt;ref name=&quot;ket&quot;&gt;https://www.ncbi.nlm.nih.gov/pubmed/27037069/&lt;/ref&gt;. Those cells that undergo cell death lose their adherence, which shows as reducing amount of CV staining in a culture &lt;ref name=&quot;ket&quot; /&gt;. Absorbance (A600) measurement after a 24 h treatment period had shown that T7DspB phage removes biofilms with more efficiency than wild-type and especially T7Control. To confirm primary results, additional test such as sonication were ran, to obtain viable cell counts (CFU per peg) for bacteria surviving in biofilms after treatment. The results show consistency with CV, and confirm that T7DspB shows much more promising signs of biofilm removal than other tested phages, especially in comparison with T7Control. As mentioned above, the idea is based on a two-pronged attack strategy. Promising results were obtained for enzymatic activity of DspB, but also we have to ensure that phages sustain sufficient replication. PFU counts from microtiter plate wells and biofilms (after sonication) show that PFU counts were significantly higher in plates than biofilms, and that overall PFU surpassed initial 10&lt;sup&gt;3&lt;/sup&gt; PFU by a few orders of magnitude which confirms efficient phage multiplication. <br /> <br /> == Time courses and dose-response for enzymatically active bacteriophage treatment ==<br /> <br /> Experiments to optimize the time-course and dose-response for both enzymatic activity and phage replication were carried out. Time-course results shown that after a 24h period of treatment, T7DspB had biofilms cell densities of two magnitudes lower than T7Control and 99,9% in comparison with untreated biofilms. Further on, time dependence on phage replication was tested, and showed that both T7Control and T7DspB began to multiply rapidly after inoculation in a similar fashion. Dosage response showed that T7DspB had lower cell densities at starting inoculation levels (PFU 10&lt;sup&gt;2&lt;/sup&gt;) than T7Control, while higher inoculation concentrations showed even more efficiency against removing biofilms. Phage dosage tested exhibited phage multiplication within the biofilm.<br /> <br /> == Discussion ==<br /> <br /> The following experiments have shown that enzymatically modified phage shows greater efficiency in biofilm removal than natural accruing phages. Future improvements to this design may include directed evolution for optimal enzyme activity, delaying cell lysis or using multiple phage promoters to allow for increased enzyme production, targeting multiple biofilm EPS components with different proteins as well as targeting multi-species biofilm with a mixture of different species-specific engineered enzymatically active phage, and combination therapy with antibiotics and phage to improve the efficacy of both types of treatment. This strategy allows opens a possibility of establishing a library of biofilm dispersing phage. The upside of this method is that it does not need to deliver, express and purify large enzyme concentrations to the site of infection. This type of phage therapy should be looked into as additional therapy for treating bacterial biofilms in various industries, but not before several challenges are overcome. Firstly, a properly designed clinical trial is needed, which will tackle the problems such as phage development resistance, immunogenicity in humans, body clearance, release of toxins after cell lysis and phage specificity. Once all the challenges are overcome, phage therapy against biofilms will be considered as our first line of defense.</div>

Roberta Mulac