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Antea-Glyphosate: Detekcija in razgradnja glifosata - Revision history
2024-03-29T14:24:17Z
Revision history for this page on the wiki
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https://wiki.fkkt.uni-lj.si/index.php?title=Antea-Glyphosate:_Detekcija_in_razgradnja_glifosata&diff=18545&oldid=prev
JernejI at 18:46, 19 April 2021
2021-04-19T18:46:14Z
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<td colspan="2" style="background-color: #fff; color: #202122; text-align: center;">Revision as of 18:46, 19 April 2021</td>
</tr><tr><td colspan="2" class="diff-lineno" id="mw-diff-left-l23">Line 23:</td>
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<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>Da bi se izognili alternativnim metabolnim potem in lažnim rezultatom zaradi citosolnega NADPH, so encime želeli pritrditi na zunanjo površino bakterijske celice, tako da bi jih fuzirali z membranskimi proteini. Kot fuzijske partnerje so preizkusili proteine INPNC, BrkA in AIDA.</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>Da bi se izognili alternativnim metabolnim potem in lažnim rezultatom zaradi citosolnega NADPH, so encime želeli pritrditi na zunanjo površino bakterijske celice, tako da bi jih fuzirali z membranskimi proteini. Kot fuzijske partnerje so preizkusili proteine INPNC, BrkA in AIDA.</div></td></tr>
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<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>==Eksperiment==</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>==Eksperiment==</div></td></tr>
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<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>*Pripravili so suspenzijo, kjer so hkrati dodali celice z vsakim od treh proteinov (pritrjenih preko INPNC). Suspenziji so dodali glifosat in NADPH ter spremljali fluorescenco skozi čas. V primerjavi s kontrolo brez izraženih proteinov je v suspenziji prišlo do znatnega upada fluorescence, s čimer so potrdili delovanje njihovega sistema.</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>*Pripravili so suspenzijo, kjer so hkrati dodali celice z vsakim od treh proteinov (pritrjenih preko INPNC). Suspenziji so dodali glifosat in NADPH ter spremljali fluorescenco skozi čas. V primerjavi s kontrolo brez izraženih proteinov je v suspenziji prišlo do znatnega upada fluorescence, s čimer so potrdili delovanje njihovega sistema.</div></td></tr>
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<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>=Razgradnja=</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>=Razgradnja=</div></td></tr>
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<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>Encim PhnO katalizira prenos acetilne skupine iz acetil koencima A na AMPA. Acetiliran AMPA lahko deluje kot substrat za PhnGHIJK, ki ga razgradi do N-metilacetamida. PhnO in PhnGHIJK lahko torej razgrajujejo tako glifosat kot tudi AMPA [6].</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>Encim PhnO katalizira prenos acetilne skupine iz acetil koencima A na AMPA. Acetiliran AMPA lahko deluje kot substrat za PhnGHIJK, ki ga razgradi do N-metilacetamida. PhnO in PhnGHIJK lahko torej razgrajujejo tako glifosat kot tudi AMPA [6].</div></td></tr>
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<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>==Eksperimenti==</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>==Eksperimenti==</div></td></tr>
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<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>S konstruktom, ki je vseboval pripravljeno biokocko, so transformirali ''E. coli'' BL21(DE3). Bakterije so gojili v gojišču z glifosatom in po 5 h določili koncentracijo preostalega glifosata v gojišču tako, da so uporabili svoj sistem za detekcijo. Primerjali so konstrukte z različnimi kombinacijami zapisov, vendar je bila najuspešnejša biokocka opisana zgoraj, ki je vsebovala vse zapise hkrati. Medtem ko so bakterije v negativni kontroli po 5 h privzele 8,6 % glifosata iz gojišča, so ''E. coli'' z biokocko privzele 43,1 %.</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>S konstruktom, ki je vseboval pripravljeno biokocko, so transformirali ''E. coli'' BL21(DE3). Bakterije so gojili v gojišču z glifosatom in po 5 h določili koncentracijo preostalega glifosata v gojišču tako, da so uporabili svoj sistem za detekcijo. Primerjali so konstrukte z različnimi kombinacijami zapisov, vendar je bila najuspešnejša biokocka opisana zgoraj, ki je vsebovala vse zapise hkrati. Medtem ko so bakterije v negativni kontroli po 5 h privzele 8,6 % glifosata iz gojišča, so ''E. coli'' z biokocko privzele 43,1 %.</div></td></tr>
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<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>=Stikala smrti=</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>=Stikala smrti=</div></td></tr>
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<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>Pripravili so eksperiment, kjer so celice z inverterjem gojili v prisotnosti ali odsotnosti arabinoze. Rast bakterij so določili s štetjem CFU po različnih časih inkubacije. Ob prisotnosti arabinoze niso opazili znatnih sprememb v številu kolonij, medtem ko je pri odsotnosti arabinoze število kolonij izrazito upadlo v roku 8 h.</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>Pripravili so eksperiment, kjer so celice z inverterjem gojili v prisotnosti ali odsotnosti arabinoze. Rast bakterij so določili s štetjem CFU po različnih časih inkubacije. Ob prisotnosti arabinoze niso opazili znatnih sprememb v številu kolonij, medtem ko je pri odsotnosti arabinoze število kolonij izrazito upadlo v roku 8 h.</div></td></tr>
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<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>==Stikalo za sistem degradacije==</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>==Stikalo za sistem degradacije==</div></td></tr>
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<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>Zaradi omejenega časa za eksperimentalno delo, so utegnili preizkusiti le delovanje posameznih sestavnih delov tega stikala, niso pa uspeli izvesti eksperimenta s celotno biokocko.</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>Zaradi omejenega časa za eksperimentalno delo, so utegnili preizkusiti le delovanje posameznih sestavnih delov tega stikala, niso pa uspeli izvesti eksperimenta s celotno biokocko.</div></td></tr>
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JernejI
https://wiki.fkkt.uni-lj.si/index.php?title=Antea-Glyphosate:_Detekcija_in_razgradnja_glifosata&diff=18544&oldid=prev
JernejI: /* Eksperimenti */
2021-04-19T18:43:25Z
<p><span dir="auto"><span class="autocomment">Eksperimenti</span></span></p>
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<td colspan="2" style="background-color: #fff; color: #202122; text-align: center;">← Older revision</td>
<td colspan="2" style="background-color: #fff; color: #202122; text-align: center;">Revision as of 18:43, 19 April 2021</td>
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<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td></tr>
<tr><td class="diff-marker" data-marker="−"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #ffe49c; vertical-align: top; white-space: pre-wrap;"><div>Da bi izboljšali razgradnjo glifosata in AMPA v ''E. coli'' so želeli nekatere gene ''phn'' nadomestiti s homologi iz drugih organizmov, kjer ta razgradnja poteka učinkoviteje. Vstaviti so želeli gene ''phnJ'' iz ''Enterobacter cloacae'' K7 [7], ''phnO'' iz ''Salmonella enterica'' [8] in ''phnE1/E2'' iz ''Sinorhizobium meliloti'' 1021 [9]. Hkrati so želeli inhibirati izražanje endognih zapisov za PhnF, ki deluje kot represor za nekatere gene ''phn'' [6], in PhnJ, ki bi lahko z rekombninantnim PhnJ tekmoval za podenote kompleksa PhnGHJIK. Kot metodo inhibicije izražanja so izbrali siRNA. </div></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div>Da bi izboljšali razgradnjo glifosata in AMPA v ''E. coli'' so želeli nekatere gene ''phn'' nadomestiti s homologi iz drugih organizmov, kjer ta razgradnja poteka učinkoviteje <ins style="font-weight: bold; text-decoration: none;">(tabela 2)</ins>. Vstaviti so želeli gene ''phnJ'' iz ''Enterobacter cloacae'' K7 [7], ''phnO'' iz ''Salmonella enterica'' [8] in ''phnE1/E2'' iz ''Sinorhizobium meliloti'' 1021 [9]. Hkrati so želeli inhibirati izražanje endognih zapisov za PhnF, ki deluje kot represor za nekatere gene ''phn'' [6], in PhnJ, ki bi lahko z rekombninantnim PhnJ tekmoval za podenote kompleksa PhnGHJIK. Kot metodo inhibicije izražanja so izbrali siRNA. </div></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>Pripravili so biokocko [http://parts.igem.org/Part:BBa_K3332099 BBa_K3332099], ki je vsebovala vse potrebne zapise. Izražanje genov za proteine so nadzorovali promotorji [http://parts.igem.org/Part:BBa_J23100 J23100] in RBS [http://parts.igem.org/Part:BBa_B0034 BBa_B0034], tako da sta imela ''phnJ'' in ''phnO'' vsak svoj promotor in RBS, ''phnE1'' in ''phnE2'' pa sta bila povezana policistronsko. Uporabili so terminator [http://parts.igem.org/Part:BBa_B0015 BBa_B0015]. Za siRNA so uporabili promotor [http://parts.igem.org/Part:BBa_J23101 J23101] in terminator [http://parts.igem.org/Part:BBa_J61048 BBa_J61048], zaporedjem pa so za boljšo stabilnost in učinkovitost dodali še zaporedje OmpA 5' UTR [10] in vezavno mesto za Hfq [11].</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>Pripravili so biokocko [http://parts.igem.org/Part:BBa_K3332099 BBa_K3332099], ki je vsebovala vse potrebne zapise. Izražanje genov za proteine so nadzorovali promotorji [http://parts.igem.org/Part:BBa_J23100 J23100] in RBS [http://parts.igem.org/Part:BBa_B0034 BBa_B0034], tako da sta imela ''phnJ'' in ''phnO'' vsak svoj promotor in RBS, ''phnE1'' in ''phnE2'' pa sta bila povezana policistronsko. Uporabili so terminator [http://parts.igem.org/Part:BBa_B0015 BBa_B0015]. Za siRNA so uporabili promotor [http://parts.igem.org/Part:BBa_J23101 J23101] in terminator [http://parts.igem.org/Part:BBa_J61048 BBa_J61048], zaporedjem pa so za boljšo stabilnost in učinkovitost dodali še zaporedje OmpA 5' UTR [10] in vezavno mesto za Hfq [11].</div></td></tr>
</table>
JernejI
https://wiki.fkkt.uni-lj.si/index.php?title=Antea-Glyphosate:_Detekcija_in_razgradnja_glifosata&diff=18543&oldid=prev
JernejI at 18:42, 19 April 2021
2021-04-19T18:42:54Z
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<td colspan="2" style="background-color: #fff; color: #202122; text-align: center;">Revision as of 18:42, 19 April 2021</td>
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<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>==Eksperimenti==</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>==Eksperimenti==</div></td></tr>
<tr><td colspan="2" class="diff-side-deleted"></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;"></ins></div></td></tr>
<tr><td colspan="2" class="diff-side-deleted"></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;">Tabela 2: Sestavni deli biokocke za degradacijo.</ins></div></td></tr>
<tr><td colspan="2" class="diff-side-deleted"></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;">{| border=1 style="border-collapse: collapse;"</ins></div></td></tr>
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<tr><td colspan="2" class="diff-side-deleted"></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;">| Zapis || Opis</ins></div></td></tr>
<tr><td colspan="2" class="diff-side-deleted"></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;">|-</ins></div></td></tr>
<tr><td colspan="2" class="diff-side-deleted"></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;">|'''PhnJ'''|| Katalitična podenota C-P liaze iz ''Enterobacter cloacae'' K7.</ins></div></td></tr>
<tr><td colspan="2" class="diff-side-deleted"></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;">|-</ins></div></td></tr>
<tr><td colspan="2" class="diff-side-deleted"></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;">|'''PhnO''' || Encim za acetilacijo AMPA iz ''Salmonella enterica''.</ins></div></td></tr>
<tr><td colspan="2" class="diff-side-deleted"></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;">|-</ins></div></td></tr>
<tr><td colspan="2" class="diff-side-deleted"></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;">|'''PhnE1 in PhnE2''' || Del transportnega sistema za glifosat iz iz ''Sinorhizobium meliloti'' 1021.</ins></div></td></tr>
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<tr><td colspan="2" class="diff-side-deleted"></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;">|'''anti-PhnJ''' || siRNA za endogen ''phnJ''.</ins></div></td></tr>
<tr><td colspan="2" class="diff-side-deleted"></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;">|-</ins></div></td></tr>
<tr><td colspan="2" class="diff-side-deleted"></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;">|'''anti-PhnF''' || siRNA za endogen ''phnF''.</ins></div></td></tr>
<tr><td colspan="2" class="diff-side-deleted"></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;">|-</ins></div></td></tr>
<tr><td colspan="2" class="diff-side-deleted"></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;">|}</ins></div></td></tr>
<tr><td colspan="2" class="diff-side-deleted"></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;"></ins></div></td></tr>
<tr><td colspan="2" class="diff-side-deleted"></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;"></ins></div></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>Da bi izboljšali razgradnjo glifosata in AMPA v ''E. coli'' so želeli nekatere gene ''phn'' nadomestiti s homologi iz drugih organizmov, kjer ta razgradnja poteka učinkoviteje. Vstaviti so želeli gene ''phnJ'' iz ''Enterobacter cloacae'' K7 [7], ''phnO'' iz ''Salmonella enterica'' [8] in ''phnE1/E2'' iz ''Sinorhizobium meliloti'' 1021 [9]. Hkrati so želeli inhibirati izražanje endognih zapisov za PhnF, ki deluje kot represor za nekatere gene ''phn'' [6], in PhnJ, ki bi lahko z rekombninantnim PhnJ tekmoval za podenote kompleksa PhnGHJIK. Kot metodo inhibicije izražanja so izbrali siRNA. </div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>Da bi izboljšali razgradnjo glifosata in AMPA v ''E. coli'' so želeli nekatere gene ''phn'' nadomestiti s homologi iz drugih organizmov, kjer ta razgradnja poteka učinkoviteje. Vstaviti so želeli gene ''phnJ'' iz ''Enterobacter cloacae'' K7 [7], ''phnO'' iz ''Salmonella enterica'' [8] in ''phnE1/E2'' iz ''Sinorhizobium meliloti'' 1021 [9]. Hkrati so želeli inhibirati izražanje endognih zapisov za PhnF, ki deluje kot represor za nekatere gene ''phn'' [6], in PhnJ, ki bi lahko z rekombninantnim PhnJ tekmoval za podenote kompleksa PhnGHJIK. Kot metodo inhibicije izražanja so izbrali siRNA. </div></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td></tr>
<tr><td class="diff-marker" data-marker="−"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #ffe49c; vertical-align: top; white-space: pre-wrap;"><div>Pripravili so biokocko BBa_K3332099, ki je vsebovala vse potrebne zapise. Izražanje genov za proteine so nadzorovali promotorji J23100 in RBS BBa_B0034, tako da sta imela ''phnJ'' in ''phnO'' vsak svoj promotor in RBS, ''phnE1'' in ''phnE2'' pa sta bila povezana policistronsko. Uporabili so terminator BBa_B0015. Za siRNA so uporabili promotor BBa_J23101 <del style="font-weight: bold; text-decoration: none;"> </del>in terminator BBa_J61048, zaporedjem pa so za boljšo stabilnost in učinkovitost dodali še zaporedje OmpA 5' UTR [10] in vezavno mesto za Hfq [11].</div></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div>Pripravili so biokocko <ins style="font-weight: bold; text-decoration: none;">[http://parts.igem.org/Part:BBa_K3332099 </ins>BBa_K3332099<ins style="font-weight: bold; text-decoration: none;">]</ins>, ki je vsebovala vse potrebne zapise. Izražanje genov za proteine so nadzorovali promotorji <ins style="font-weight: bold; text-decoration: none;">[http://parts.igem.org/Part:BBa_J23100 </ins>J23100<ins style="font-weight: bold; text-decoration: none;">] </ins>in RBS <ins style="font-weight: bold; text-decoration: none;">[http://parts.igem.org/Part:BBa_B0034 </ins>BBa_B0034<ins style="font-weight: bold; text-decoration: none;">]</ins>, tako da sta imela ''phnJ'' in ''phnO'' vsak svoj promotor in RBS, ''phnE1'' in ''phnE2'' pa sta bila povezana policistronsko. Uporabili so terminator <ins style="font-weight: bold; text-decoration: none;">[http://parts.igem.org/Part:BBa_B0015 </ins>BBa_B0015<ins style="font-weight: bold; text-decoration: none;">]</ins>. Za siRNA so uporabili promotor <ins style="font-weight: bold; text-decoration: none;">[http://parts.igem.org/Part:</ins>BBa_J23101 <ins style="font-weight: bold; text-decoration: none;">J23101] </ins>in terminator <ins style="font-weight: bold; text-decoration: none;">[http://parts.igem.org/Part:BBa_J61048 </ins>BBa_J61048<ins style="font-weight: bold; text-decoration: none;">]</ins>, zaporedjem pa so za boljšo stabilnost in učinkovitost dodali še zaporedje OmpA 5' UTR [10] in vezavno mesto za Hfq [11].</div></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>S konstruktom, ki je vseboval pripravljeno biokocko, so transformirali ''E. coli'' BL21(DE3). Bakterije so gojili v gojišču z glifosatom in po 5 h določili koncentracijo preostalega glifosata v gojišču tako, da so uporabili svoj sistem za detekcijo. Primerjali so konstrukte z različnimi kombinacijami zapisov, vendar je bila najuspešnejša biokocka opisana zgoraj, ki je vsebovala vse zapise hkrati. Medtem ko so bakterije v negativni kontroli po 5 h privzele 8,6 % glifosata iz gojišča, so ''E. coli'' z biokocko privzele 43,1 %.</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>S konstruktom, ki je vseboval pripravljeno biokocko, so transformirali ''E. coli'' BL21(DE3). Bakterije so gojili v gojišču z glifosatom in po 5 h določili koncentracijo preostalega glifosata v gojišču tako, da so uporabili svoj sistem za detekcijo. Primerjali so konstrukte z različnimi kombinacijami zapisov, vendar je bila najuspešnejša biokocka opisana zgoraj, ki je vsebovala vse zapise hkrati. Medtem ko so bakterije v negativni kontroli po 5 h privzele 8,6 % glifosata iz gojišča, so ''E. coli'' z biokocko privzele 43,1 %.</div></td></tr>
</table>
JernejI
https://wiki.fkkt.uni-lj.si/index.php?title=Antea-Glyphosate:_Detekcija_in_razgradnja_glifosata&diff=18541&oldid=prev
JernejI: /* Stikalo za sistem detekcije */
2021-04-19T18:29:24Z
<p><span dir="auto"><span class="autocomment">Stikalo za sistem detekcije</span></span></p>
<table style="background-color: #fff; color: #202122;" data-mw="interface">
<col class="diff-marker" />
<col class="diff-content" />
<col class="diff-marker" />
<col class="diff-content" />
<tr class="diff-title" lang="en">
<td colspan="2" style="background-color: #fff; color: #202122; text-align: center;">← Older revision</td>
<td colspan="2" style="background-color: #fff; color: #202122; text-align: center;">Revision as of 18:29, 19 April 2021</td>
</tr><tr><td colspan="2" class="diff-lineno" id="mw-diff-left-l69">Line 69:</td>
<td colspan="2" class="diff-lineno">Line 69:</td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>Stikalo smrti pri sistemu detekcije glifosata bi zagotovilo, da bi gensko spremenjene ''E. coli'' odmrle, če bi se sprostile v okolje. Gojišče, v katerem bi potekala detekcija glifosata v vzorcu, bi vsebovalo arabinozo, ki bi inhibirala sintezo citotoksične nukleaze MazF [12]. V primeru, da bi bakterije zapustile to goijšče, bi se mazF lahko izražal in privedel do celične smrti.</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>Stikalo smrti pri sistemu detekcije glifosata bi zagotovilo, da bi gensko spremenjene ''E. coli'' odmrle, če bi se sprostile v okolje. Gojišče, v katerem bi potekala detekcija glifosata v vzorcu, bi vsebovalo arabinozo, ki bi inhibirala sintezo citotoksične nukleaze MazF [12]. V primeru, da bi bakterije zapustile to goijšče, bi se mazF lahko izražal in privedel do celične smrti.</div></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td></tr>
<tr><td class="diff-marker" data-marker="−"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #ffe49c; vertical-align: top; white-space: pre-wrap;"><div>Pripravili so preprosto napravo [http://parts.igem.org/Part:<del style="font-weight: bold; text-decoration: none;">BBa_K3332078 BBa_K3332078</del>], ki deluje kot inverter. Promotor P<sub>BAD</sub> nadzoruje izražanje zapisa za cI, promotor P<sub>R</sub> pa nadzoruje izražanje ''mazF''. Arabinoza povzroči preklop araC iz represorske v aktivatorsko obliko, kar spodbudi izražanje zapisa za cI, cI pa inhibira transkripcijo ''mazF''. V odsotnosti arabinoze bo araC inhibiral izražanje ''cI'', torej bo transkripcija ''mazF'' lahko potekala.</div></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div>Pripravili so preprosto napravo [http://parts.igem.org/Part:<ins style="font-weight: bold; text-decoration: none;">BBa_K3332081 BBa_K3332081</ins>], ki deluje kot inverter. Promotor P<sub>BAD</sub> nadzoruje izražanje zapisa za cI, promotor P<sub>R</sub> pa nadzoruje izražanje ''mazF''. Arabinoza povzroči preklop araC iz represorske v aktivatorsko obliko, kar spodbudi izražanje zapisa za cI, cI pa inhibira transkripcijo ''mazF''. V odsotnosti arabinoze bo araC inhibiral izražanje ''cI'', torej bo transkripcija ''mazF'' lahko potekala.</div></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>Pripravili so eksperiment, kjer so celice z inverterjem gojili v prisotnosti ali odsotnosti arabinoze. Rast bakterij so določili s štetjem CFU po različnih časih inkubacije. Ob prisotnosti arabinoze niso opazili znatnih sprememb v številu kolonij, medtem ko je pri odsotnosti arabinoze število kolonij izrazito upadlo v roku 8 h.</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>Pripravili so eksperiment, kjer so celice z inverterjem gojili v prisotnosti ali odsotnosti arabinoze. Rast bakterij so določili s štetjem CFU po različnih časih inkubacije. Ob prisotnosti arabinoze niso opazili znatnih sprememb v številu kolonij, medtem ko je pri odsotnosti arabinoze število kolonij izrazito upadlo v roku 8 h.</div></td></tr>
</table>
JernejI
https://wiki.fkkt.uni-lj.si/index.php?title=Antea-Glyphosate:_Detekcija_in_razgradnja_glifosata&diff=18540&oldid=prev
JernejI: /* Stikalo za sistem degradacije */
2021-04-19T18:28:24Z
<p><span dir="auto"><span class="autocomment">Stikalo za sistem degradacije</span></span></p>
<table style="background-color: #fff; color: #202122;" data-mw="interface">
<col class="diff-marker" />
<col class="diff-content" />
<col class="diff-marker" />
<col class="diff-content" />
<tr class="diff-title" lang="en">
<td colspan="2" style="background-color: #fff; color: #202122; text-align: center;">← Older revision</td>
<td colspan="2" style="background-color: #fff; color: #202122; text-align: center;">Revision as of 18:28, 19 April 2021</td>
</tr><tr><td colspan="2" class="diff-lineno" id="mw-diff-left-l77">Line 77:</td>
<td colspan="2" class="diff-lineno">Line 77:</td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>Zamislili so si napravo, ki bi ob prisotnosti glifosata zavirala izražanje ''mazF'', v njegovi odsotnosti pa se bi ''mazF'' izražal in vodil v smrt.</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>Zamislili so si napravo, ki bi ob prisotnosti glifosata zavirala izražanje ''mazF'', v njegovi odsotnosti pa se bi ''mazF'' izražal in vodil v smrt.</div></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td></tr>
<tr><td class="diff-marker" data-marker="−"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #ffe49c; vertical-align: top; white-space: pre-wrap;"><div>Biokocka BBa_K3332078 deluje kot inverter. Namesto neposredne detekcije glifosata, so kot merilo njegove prisotnosti uporabili formaldehid, ki je eden od produktov ob razgradnji glifosata. HxlR je transkripcijski faktor, ki ob vezavi formaldehida deluje kot aktivator promotorja P<sub>HCHO</sub> [13]. P<sub>HCHO</sub> nadzoruje izražanje ''cI'', promotor P<sub>R</sub> pa izražanje ''mazF''. Ob prisotnosti formaldehida, bo HxlR aktiviral transkripcijo zapisa za cI, cI pa bo onemogočil izražanje ''mazF''. Kadar pa je formaldehid odsoten, transkripcija ''cI'' ne bo potekala in posledično se ''mazF'' lahko izraža.</div></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div>Biokocka <ins style="font-weight: bold; text-decoration: none;">[http://parts.igem.org/Part:</ins>BBa_K3332078 <ins style="font-weight: bold; text-decoration: none;">BBa_K3332078] </ins>deluje kot inverter. Namesto neposredne detekcije glifosata, so kot merilo njegove prisotnosti uporabili formaldehid, ki je eden od produktov ob razgradnji glifosata. HxlR je transkripcijski faktor, ki ob vezavi formaldehida deluje kot aktivator promotorja P<sub>HCHO</sub> [13]. P<sub>HCHO</sub> nadzoruje izražanje ''cI'', promotor P<sub>R</sub> pa izražanje ''mazF''. Ob prisotnosti formaldehida, bo HxlR aktiviral transkripcijo zapisa za cI, cI pa bo onemogočil izražanje ''mazF''. Kadar pa je formaldehid odsoten, transkripcija ''cI'' ne bo potekala in posledično se ''mazF'' lahko izraža.</div></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>Zaradi omejenega časa za eksperimentalno delo, so utegnili preizkusiti le delovanje posameznih sestavnih delov tega stikala, niso pa uspeli izvesti eksperimenta s celotno biokocko.</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>Zaradi omejenega časa za eksperimentalno delo, so utegnili preizkusiti le delovanje posameznih sestavnih delov tega stikala, niso pa uspeli izvesti eksperimenta s celotno biokocko.</div></td></tr>
</table>
JernejI
https://wiki.fkkt.uni-lj.si/index.php?title=Antea-Glyphosate:_Detekcija_in_razgradnja_glifosata&diff=18539&oldid=prev
JernejI: /* Stikalo za sistem detekcije */
2021-04-19T18:28:00Z
<p><span dir="auto"><span class="autocomment">Stikalo za sistem detekcije</span></span></p>
<table style="background-color: #fff; color: #202122;" data-mw="interface">
<col class="diff-marker" />
<col class="diff-content" />
<col class="diff-marker" />
<col class="diff-content" />
<tr class="diff-title" lang="en">
<td colspan="2" style="background-color: #fff; color: #202122; text-align: center;">← Older revision</td>
<td colspan="2" style="background-color: #fff; color: #202122; text-align: center;">Revision as of 18:28, 19 April 2021</td>
</tr><tr><td colspan="2" class="diff-lineno" id="mw-diff-left-l69">Line 69:</td>
<td colspan="2" class="diff-lineno">Line 69:</td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>Stikalo smrti pri sistemu detekcije glifosata bi zagotovilo, da bi gensko spremenjene ''E. coli'' odmrle, če bi se sprostile v okolje. Gojišče, v katerem bi potekala detekcija glifosata v vzorcu, bi vsebovalo arabinozo, ki bi inhibirala sintezo citotoksične nukleaze MazF [12]. V primeru, da bi bakterije zapustile to goijšče, bi se mazF lahko izražal in privedel do celične smrti.</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>Stikalo smrti pri sistemu detekcije glifosata bi zagotovilo, da bi gensko spremenjene ''E. coli'' odmrle, če bi se sprostile v okolje. Gojišče, v katerem bi potekala detekcija glifosata v vzorcu, bi vsebovalo arabinozo, ki bi inhibirala sintezo citotoksične nukleaze MazF [12]. V primeru, da bi bakterije zapustile to goijšče, bi se mazF lahko izražal in privedel do celične smrti.</div></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td></tr>
<tr><td class="diff-marker" data-marker="−"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #ffe49c; vertical-align: top; white-space: pre-wrap;"><div>Pripravili so preprosto napravo BBa_K3332078, ki deluje kot inverter. Promotor P<sub>BAD</sub> nadzoruje izražanje zapisa za cI, promotor P<sub>R</sub> pa nadzoruje izražanje ''mazF''. Arabinoza povzroči preklop araC iz represorske v aktivatorsko obliko, kar spodbudi izražanje zapisa za cI, cI pa inhibira transkripcijo ''mazF''. V odsotnosti arabinoze bo araC inhibiral izražanje ''cI'', torej bo transkripcija ''mazF'' lahko potekala.</div></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div>Pripravili so preprosto napravo <ins style="font-weight: bold; text-decoration: none;">[http://parts.igem.org/Part:</ins>BBa_K3332078 <ins style="font-weight: bold; text-decoration: none;">BBa_K3332078]</ins>, ki deluje kot inverter. Promotor P<sub>BAD</sub> nadzoruje izražanje zapisa za cI, promotor P<sub>R</sub> pa nadzoruje izražanje ''mazF''. Arabinoza povzroči preklop araC iz represorske v aktivatorsko obliko, kar spodbudi izražanje zapisa za cI, cI pa inhibira transkripcijo ''mazF''. V odsotnosti arabinoze bo araC inhibiral izražanje ''cI'', torej bo transkripcija ''mazF'' lahko potekala.</div></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>Pripravili so eksperiment, kjer so celice z inverterjem gojili v prisotnosti ali odsotnosti arabinoze. Rast bakterij so določili s štetjem CFU po različnih časih inkubacije. Ob prisotnosti arabinoze niso opazili znatnih sprememb v številu kolonij, medtem ko je pri odsotnosti arabinoze število kolonij izrazito upadlo v roku 8 h.</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>Pripravili so eksperiment, kjer so celice z inverterjem gojili v prisotnosti ali odsotnosti arabinoze. Rast bakterij so določili s štetjem CFU po različnih časih inkubacije. Ob prisotnosti arabinoze niso opazili znatnih sprememb v številu kolonij, medtem ko je pri odsotnosti arabinoze število kolonij izrazito upadlo v roku 8 h.</div></td></tr>
</table>
JernejI
https://wiki.fkkt.uni-lj.si/index.php?title=Antea-Glyphosate:_Detekcija_in_razgradnja_glifosata&diff=18537&oldid=prev
JernejI: /* Eksperiment */
2021-04-19T18:25:32Z
<p><span dir="auto"><span class="autocomment">Eksperiment</span></span></p>
<table style="background-color: #fff; color: #202122;" data-mw="interface">
<col class="diff-marker" />
<col class="diff-content" />
<col class="diff-marker" />
<col class="diff-content" />
<tr class="diff-title" lang="en">
<td colspan="2" style="background-color: #fff; color: #202122; text-align: center;">← Older revision</td>
<td colspan="2" style="background-color: #fff; color: #202122; text-align: center;">Revision as of 18:25, 19 April 2021</td>
</tr><tr><td colspan="2" class="diff-lineno" id="mw-diff-left-l26">Line 26:</td>
<td colspan="2" class="diff-lineno">Line 26:</td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>==Eksperiment==</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>==Eksperiment==</div></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td></tr>
<tr><td class="diff-marker" data-marker="−"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #ffe49c; vertical-align: top; white-space: pre-wrap;"><div>Tabela 1: <del style="font-weight: bold; text-decoration: none;">Povzetek pripravljenih biokock </del>za <del style="font-weight: bold; text-decoration: none;">razgradnjo</del></div></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div>Tabela 1: <ins style="font-weight: bold; text-decoration: none;">Biokocke </ins>za <ins style="font-weight: bold; text-decoration: none;">sistem detekcije.</ins></div></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>{| border=1 style="border-collapse: collapse;"</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>{| border=1 style="border-collapse: collapse;"</div></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>|-</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>|-</div></td></tr>
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<td colspan="2" class="diff-lineno">Line 38:</td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>|-</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>|-</div></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>|}</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>|}</div></td></tr>
<tr><td colspan="2" class="diff-side-deleted"></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;"></ins></div></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>V eksperimentu so želeli preizkusiti delovanje proteinov, ko so bili ti pritrjeni na površino celic. V predhodnih eksperimentih so potrdili uspešno izražanje izbranih proteinov na površini celice. Pripravili so biokocke, kjer je bil vsak izmed treh proteinov fuziran na tri različne membranske proteine: INPNC, BrkA ali AIDA (tabela 1). Biokocke so vsebovale konstitutivni promotor [http://parts.igem.org/Part:BBa_J23100 J23100], RBS [http://parts.igem.org/Part:BBa_B0034 BBa_B0034] in terminator [http://parts.igem.org/Part:BBa_B0015 BBa_B0015]. Proteine so predstavili na površini ''E. coli'' DH5α in jih preizkusili v seriji eksperimentov.</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>V eksperimentu so želeli preizkusiti delovanje proteinov, ko so bili ti pritrjeni na površino celic. V predhodnih eksperimentih so potrdili uspešno izražanje izbranih proteinov na površini celice. Pripravili so biokocke, kjer je bil vsak izmed treh proteinov fuziran na tri različne membranske proteine: INPNC, BrkA ali AIDA (tabela 1). Biokocke so vsebovale konstitutivni promotor [http://parts.igem.org/Part:BBa_J23100 J23100], RBS [http://parts.igem.org/Part:BBa_B0034 BBa_B0034] in terminator [http://parts.igem.org/Part:BBa_B0015 BBa_B0015]. Proteine so predstavili na površini ''E. coli'' DH5α in jih preizkusili v seriji eksperimentov.</div></td></tr>
</table>
JernejI
https://wiki.fkkt.uni-lj.si/index.php?title=Antea-Glyphosate:_Detekcija_in_razgradnja_glifosata&diff=18535&oldid=prev
JernejI: /* Eksperiment */
2021-04-19T18:24:07Z
<p><span dir="auto"><span class="autocomment">Eksperiment</span></span></p>
<table style="background-color: #fff; color: #202122;" data-mw="interface">
<col class="diff-marker" />
<col class="diff-content" />
<col class="diff-marker" />
<col class="diff-content" />
<tr class="diff-title" lang="en">
<td colspan="2" style="background-color: #fff; color: #202122; text-align: center;">← Older revision</td>
<td colspan="2" style="background-color: #fff; color: #202122; text-align: center;">Revision as of 18:24, 19 April 2021</td>
</tr><tr><td colspan="2" class="diff-lineno" id="mw-diff-left-l31">Line 31:</td>
<td colspan="2" class="diff-lineno">Line 31:</td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>| Protein || Opis || '''INPNC''' || '''BrkA''' || '''AIDA'''</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>| Protein || Opis || '''INPNC''' || '''BrkA''' || '''AIDA'''</div></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>|-</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>|-</div></td></tr>
<tr><td class="diff-marker" data-marker="−"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #ffe49c; vertical-align: top; white-space: pre-wrap;"><div>| ''' GOX ''' || Glifosat oksidoreduktaza iz rastline ''Echinochloa colona''. || [http://parts.igem.org/Part:BBa_K3332052 BBa_K3332052] || [http://parts.igem.org/Part:BBa_K3332055 BBa_K3332055] || [http://parts.igem.org/Part:BBa_K3332054 BBa_K3332054]</div></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div>|''' GOX ''' || Glifosat oksidoreduktaza iz rastline ''Echinochloa colona''. || [http://parts.igem.org/Part:BBa_K3332052 BBa_K3332052] || [http://parts.igem.org/Part:BBa_K3332055 BBa_K3332055] || [http://parts.igem.org/Part:BBa_K3332054 BBa_K3332054]</div></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>|-</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>|-</div></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>| ''' GRHPR '''|| Glioksilat reduktaza iz človeških jeter. || [http://parts.igem.org/Part:BBa_K3332057 BBa_K3332057] || [http://parts.igem.org/Part:BBa_K3332060 BBa_K3332060] || [http://parts.igem.org/Part:BBa_K3332059 BBa_K3332059]</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>| ''' GRHPR '''|| Glioksilat reduktaza iz človeških jeter. || [http://parts.igem.org/Part:BBa_K3332057 BBa_K3332057] || [http://parts.igem.org/Part:BBa_K3332060 BBa_K3332060] || [http://parts.igem.org/Part:BBa_K3332059 BBa_K3332059]</div></td></tr>
</table>
JernejI
https://wiki.fkkt.uni-lj.si/index.php?title=Antea-Glyphosate:_Detekcija_in_razgradnja_glifosata&diff=18534&oldid=prev
JernejI at 18:14, 19 April 2021
2021-04-19T18:14:32Z
<p></p>
<table style="background-color: #fff; color: #202122;" data-mw="interface">
<col class="diff-marker" />
<col class="diff-content" />
<col class="diff-marker" />
<col class="diff-content" />
<tr class="diff-title" lang="en">
<td colspan="2" style="background-color: #fff; color: #202122; text-align: center;">← Older revision</td>
<td colspan="2" style="background-color: #fff; color: #202122; text-align: center;">Revision as of 18:14, 19 April 2021</td>
</tr><tr><td colspan="2" class="diff-lineno" id="mw-diff-left-l29">Line 29:</td>
<td colspan="2" class="diff-lineno">Line 29:</td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>{| border=1 style="border-collapse: collapse;"</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>{| border=1 style="border-collapse: collapse;"</div></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>|-</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>|-</div></td></tr>
<tr><td class="diff-marker" data-marker="−"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #ffe49c; vertical-align: top; white-space: pre-wrap;"><div>| Protein || Opis || INPNC || BrkA || AIDA</div></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div>| Protein || Opis || <ins style="font-weight: bold; text-decoration: none;">'''</ins>INPNC<ins style="font-weight: bold; text-decoration: none;">''' </ins>|| <ins style="font-weight: bold; text-decoration: none;">'''</ins>BrkA<ins style="font-weight: bold; text-decoration: none;">''' </ins>|| <ins style="font-weight: bold; text-decoration: none;">'''</ins>AIDA<ins style="font-weight: bold; text-decoration: none;">'''</ins></div></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>|-</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>|-</div></td></tr>
<tr><td class="diff-marker" data-marker="−"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #ffe49c; vertical-align: top; white-space: pre-wrap;"><div>| GOX || <del style="font-weight: bold; text-decoration: none;">opls </del>|| [http://parts.igem.org/Part:BBa_K3332052 BBa_K3332052] || [http://parts.igem.org/Part:BBa_K3332055 BBa_K3332055] || [http://parts.igem.org/Part:BBa_K3332054 BBa_K3332054]</div></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div>| <ins style="font-weight: bold; text-decoration: none;">''' </ins>GOX <ins style="font-weight: bold; text-decoration: none;">''' </ins>|| <ins style="font-weight: bold; text-decoration: none;">Glifosat oksidoreduktaza iz rastline ''Echinochloa colona''. </ins>|| [http://parts.igem.org/Part:BBa_K3332052 BBa_K3332052] || [http://parts.igem.org/Part:BBa_K3332055 BBa_K3332055] || [http://parts.igem.org/Part:BBa_K3332054 BBa_K3332054]</div></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>|-</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>|-</div></td></tr>
<tr><td class="diff-marker" data-marker="−"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #ffe49c; vertical-align: top; white-space: pre-wrap;"><div>| GRHPR || <del style="font-weight: bold; text-decoration: none;">opls </del>|| [http://parts.igem.org/Part:BBa_K3332057 BBa_K3332057] || [http://parts.igem.org/Part:BBa_K3332060 BBa_K3332060] || [http://parts.igem.org/Part:BBa_K3332059 BBa_K3332059]</div></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div>| <ins style="font-weight: bold; text-decoration: none;">''' </ins>GRHPR <ins style="font-weight: bold; text-decoration: none;">'''</ins>|| <ins style="font-weight: bold; text-decoration: none;">Glioksilat reduktaza iz človeških jeter. </ins>|| [http://parts.igem.org/Part:BBa_K3332057 BBa_K3332057] || [http://parts.igem.org/Part:BBa_K3332060 BBa_K3332060] || [http://parts.igem.org/Part:BBa_K3332059 BBa_K3332059]</div></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>|-</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>|-</div></td></tr>
<tr><td class="diff-marker" data-marker="−"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #ffe49c; vertical-align: top; white-space: pre-wrap;"><div>| iNAP || <del style="font-weight: bold; text-decoration: none;">opis </del>|| [http://parts.igem.org/Part:BBa_K3332047 BBa_K3332047] || [http://parts.igem.org/Part:BBa_K3332050 BBa_K3332050] || [http://parts.igem.org/Part:BBa_K3332049 BBa_K3332049]</div></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div>| <ins style="font-weight: bold; text-decoration: none;">''' </ins>iNAP <ins style="font-weight: bold; text-decoration: none;">'''</ins>|| <ins style="font-weight: bold; text-decoration: none;">Fuzija proteinov cpYFP in NADPH vezavne domene proteina REX iz ''Thermus aquaticus''.</ins>|| [http://parts.igem.org/Part:BBa_K3332047 BBa_K3332047] || [http://parts.igem.org/Part:BBa_K3332050 BBa_K3332050] || [http://parts.igem.org/Part:BBa_K3332049 BBa_K3332049]</div></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>|-</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>|-</div></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>|}</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>|}</div></td></tr>
</table>
JernejI
https://wiki.fkkt.uni-lj.si/index.php?title=Antea-Glyphosate:_Detekcija_in_razgradnja_glifosata&diff=18532&oldid=prev
JernejI: /* Eksperiment */
2021-04-19T18:05:48Z
<p><span dir="auto"><span class="autocomment">Eksperiment</span></span></p>
<table style="background-color: #fff; color: #202122;" data-mw="interface">
<col class="diff-marker" />
<col class="diff-content" />
<col class="diff-marker" />
<col class="diff-content" />
<tr class="diff-title" lang="en">
<td colspan="2" style="background-color: #fff; color: #202122; text-align: center;">← Older revision</td>
<td colspan="2" style="background-color: #fff; color: #202122; text-align: center;">Revision as of 18:05, 19 April 2021</td>
</tr><tr><td colspan="2" class="diff-lineno" id="mw-diff-left-l25">Line 25:</td>
<td colspan="2" class="diff-lineno">Line 25:</td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>==Eksperiment==</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>==Eksperiment==</div></td></tr>
<tr><td class="diff-marker" data-marker="−"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #ffe49c; vertical-align: top; white-space: pre-wrap;"><div>V eksperimentu so želeli preizkusiti delovanje proteinov, ko so bili ti pritrjeni na površino celic. V predhodnih eksperimentih so potrdili uspešno izražanje izbranih proteinov na površini celice. Pripravili so biokocke <del style="font-weight: bold; text-decoration: none;">BBa_K2922060, BBa_K3332057, BBa_K3332059 (za GRHPR), BBa_K3332052, BBa_K3332054, BBa_K3332055 (za GOX), BBa_K3332047, BBa_K3332049, BBa_K3332050 (za iNap)</del>, kjer je bil vsak izmed treh proteinov fuziran na tri različne membranske proteine: INPNC, BrkA ali AIDA. Biokocke so vsebovale promotor J23100, RBS BBa_B0034 in terminator BBa_B0015. Proteine so predstavili na površini ''E. coli'' DH5α in jih preizkusili v seriji eksperimentov.</div></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div> </div></td></tr>
<tr><td colspan="2" class="diff-side-deleted"></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;">Tabela 1: Povzetek pripravljenih biokock za razgradnjo</ins></div></td></tr>
<tr><td colspan="2" class="diff-side-deleted"></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;">{| border=1 style="border-collapse: collapse;"</ins></div></td></tr>
<tr><td colspan="2" class="diff-side-deleted"></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;">|-</ins></div></td></tr>
<tr><td colspan="2" class="diff-side-deleted"></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;">| Protein || Opis || INPNC || BrkA || AIDA</ins></div></td></tr>
<tr><td colspan="2" class="diff-side-deleted"></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;">|-</ins></div></td></tr>
<tr><td colspan="2" class="diff-side-deleted"></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;">| GOX || opls || [http://parts.igem.org/Part:BBa_K3332052 BBa_K3332052] || [http://parts.igem.org/Part:BBa_K3332055 BBa_K3332055] || [http://parts.igem.org/Part:BBa_K3332054 BBa_K3332054]</ins></div></td></tr>
<tr><td colspan="2" class="diff-side-deleted"></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;">|-</ins></div></td></tr>
<tr><td colspan="2" class="diff-side-deleted"></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;">| GRHPR || opls || [http://parts.igem.org/Part:BBa_K3332057 BBa_K3332057] || [http://parts.igem.org/Part:BBa_K3332060 BBa_K3332060] || [http://parts.igem.org/Part:BBa_K3332059 BBa_K3332059]</ins></div></td></tr>
<tr><td colspan="2" class="diff-side-deleted"></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;">|-</ins></div></td></tr>
<tr><td colspan="2" class="diff-side-deleted"></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;">| iNAP || opis || [http://parts.igem.org/Part:BBa_K3332047 BBa_K3332047] || [http://parts.igem.org/Part:BBa_K3332050 BBa_K3332050] || [http://parts.igem.org/Part:BBa_K3332049 BBa_K3332049]</ins></div></td></tr>
<tr><td colspan="2" class="diff-side-deleted"></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;">|-</ins></div></td></tr>
<tr><td colspan="2" class="diff-side-deleted"></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;">|}</ins></div></td></tr>
<tr><td colspan="2" class="diff-side-deleted"></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div> </div></td></tr>
<tr><td colspan="2" class="diff-side-deleted"></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div>V eksperimentu so želeli preizkusiti delovanje proteinov, ko so bili ti pritrjeni na površino celic. V predhodnih eksperimentih so potrdili uspešno izražanje izbranih proteinov na površini celice. Pripravili so biokocke, kjer je bil vsak izmed treh proteinov fuziran na tri različne membranske proteine: INPNC, BrkA ali AIDA <ins style="font-weight: bold; text-decoration: none;">(tabela 1)</ins>. Biokocke so vsebovale <ins style="font-weight: bold; text-decoration: none;">konstitutivni </ins>promotor <ins style="font-weight: bold; text-decoration: none;">[http://parts.igem.org/Part:BBa_J23100 </ins>J23100<ins style="font-weight: bold; text-decoration: none;">]</ins>, RBS <ins style="font-weight: bold; text-decoration: none;">[http://parts.igem.org/Part:BBa_B0034 </ins>BBa_B0034<ins style="font-weight: bold; text-decoration: none;">] </ins>in terminator <ins style="font-weight: bold; text-decoration: none;">[http://parts.igem.org/Part:BBa_B0015 </ins>BBa_B0015<ins style="font-weight: bold; text-decoration: none;">]</ins>. Proteine so predstavili na površini ''E. coli'' DH5α in jih preizkusili v seriji eksperimentov.</div></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>*Suspenziji celic z izraženim GRHPR so dodali glioksilat in NADPH. Koncentracijo NADPH so spremljali z merjenjem OD<sub>340</sub>. Opazili so, da je koncentracija NADPH skozi čas padala, na podlagi česar so sklepali, da encim uspešno katalizira reakcijo. Primerjali so učinkovitost encima z različnimi fuzijskimi partnerji in sklenili, da je bila izmed vseh najučinkovitejša kombinacija INPNC-GRHPR.</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>*Suspenziji celic z izraženim GRHPR so dodali glioksilat in NADPH. Koncentracijo NADPH so spremljali z merjenjem OD<sub>340</sub>. Opazili so, da je koncentracija NADPH skozi čas padala, na podlagi česar so sklepali, da encim uspešno katalizira reakcijo. Primerjali so učinkovitost encima z različnimi fuzijskimi partnerji in sklenili, da je bila izmed vseh najučinkovitejša kombinacija INPNC-GRHPR.</div></td></tr>
</table>
JernejI