Difference between revisions of "Part:BBa K2012023"
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<p style="text-align:justify;text-justify:inter-ideograph"><span style="font-family:Calibri"> </span></p> | <p style="text-align:justify;text-justify:inter-ideograph"><span style="font-family:Calibri"> </span></p> | ||
<p style="text-align:justify;text-justify:inter-ideograph"><span style="font-family:Calibri"><img height="293.1300048828125" src="https://static.igem.org/mediawiki/2016/7/7a/Pcpcg2.jpeg " width="480.0"></span><span style="font-family:Calibri"><span></span></span></p> | <p style="text-align:justify;text-justify:inter-ideograph"><span style="font-family:Calibri"><img height="293.1300048828125" src="https://static.igem.org/mediawiki/2016/7/7a/Pcpcg2.jpeg " width="480.0"></span><span style="font-family:Calibri"><span></span></span></p> | ||
| − | <p style="text-align:justify;text-justify:inter-ideograph"><span style="font-family:Calibri"> | + | <p style="text-align:justify;text-justify:inter-ideograph"><span style="font-family:Calibri"><b>Figure 1.</b> Comparison of sfGFP </span><span style="font-family:Calibri">fluorescence</span><span style="font-family:Calibri"> of CcaS-CcaR system under green light, red light and darkness.<span></span></span></p> |
<p style="text-align:justify;text-justify:inter-ideograph"><span style="font-family:Calibri"> </span></p> | <p style="text-align:justify;text-justify:inter-ideograph"><span style="font-family:Calibri"> </span></p> | ||
| − | <p style="text-align:justify;text-justify:inter-ideograph"><span style="font-family:Calibri">As is shown in | + | <p style="text-align:justify;text-justify:inter-ideograph"><span style="font-family:Calibri">As is shown in Figure 1, in E.coli strain JT2, PcpcG2 </span><span style="font-family:Calibri">produces </span><span style="font-family:Calibri">266.0</span><span style="font-family:Calibri"> ± 1</span><span style="font-family:Calibri">9.</span><span style="font-family:Calibri">3 and </span><span style="font-family:Calibri">509.0</span><span style="font-family:Calibri"> ± </span><span style="font-family:Calibri">55.4</span><span style="font-family:Calibri"> au of</span><span style="font-family:Calibri"> </span><span style="font-family:Calibri">sfGFP in red and green light, corresponding to </span><span style="font-family:Calibri">1.91</span><span style="font-family:Calibri"> ± 0.3</span><span style="font-family:Calibri">4</span><span style="font-family:Calibri">-fold</span><span style="font-family:Calibri"> </span><span style="font-family:Calibri">activation</span><span style="font-family:Calibri">, demonstrating that PcpcG2 is functionally regulated by light, green-activated and red-repressed.<span></span></span></p> |
<p style="text-align:justify;text-justify:inter-ideograph"><span style="font-family:Calibri"> </span></p> | <p style="text-align:justify;text-justify:inter-ideograph"><span style="font-family:Calibri"> </span></p> | ||
| − | <p style="text-align:justify;text-justify:inter-ideograph"><span style="font-family:Calibri">However, the figure also shows that that leaked expression is severe in dark surroundings. To better respond to red and green light, we optimized the promoter by refactoring it and created an innovative promoter, PcpcG2-172, with the truncation of a constitutive promoter within the cpcG2 promoter </span><span style="font-family:Calibri">(</span><span style="font-family:Calibri">BBa_K</span><span style="font-family:Calibri">2012015</span><span style="font-family:Calibri">. </span><span style="font-family:Calibri">For more detail, please view our wiki: http://2016.igem.org/Team:HZAU-China/Experiments)</span><span style="font-family:Calibri" | + | <p style="text-align:justify;text-justify:inter-ideograph"><span style="font-family:Calibri">However, the figure also shows that that leaked expression is severe in dark surroundings. To better respond to red and green light, we optimized the promoter by refactoring it and created an innovative promoter, PcpcG2-172, with the truncation of a constitutive promoter within the cpcG2 promoter </span><span style="font-family:Calibri">(</span><span style="font-family:Calibri">BBa_K</span><span style="font-family:Calibri">2012015</span><span style="font-family:Calibri">. </span><span style="font-family:Calibri">For more detail, please view our wiki: http://2016.igem.org/Team:HZAU-China/Experiments)</span><span style="font-family:Calibri"> |
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| + | <h3>Improve by HZAU-China 2017, designed by Chu Pan</h3> | ||
| + | <p>This year we improved the CcaS-CcaR system used in last year. By changing the RBS strength of CcaR, we optimized the expression of sfGFP from 2-fold variance up to 3-fold variance. For more information, please view<a href="http://2017.igem.org/Team:HZAU-China/Improve">17_HZAU-China/Improve</a> | ||
| + | <a><img src="https://parts.igem.org/File:T--HZAU-China--improve.png" width="30%"> </a> | ||
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Latest revision as of 03:27, 2 November 2017
PcpcG2-B0034-sfGFP
sfGFP Generator from pJT119b original plasmid.
Promoter cpcG2 is a 238bp green-light activated promoter from the genome of Synechocystis PCC6803. We tested the efficiency of the promoter by measuring the fluorescence of output sfGFP when bacteria are illuminated with green, red or no light. (For more detail, please view our wiki: http://2016.igem.org/Team:HZAU-China/Experiments)
Figure 1. Comparison of sfGFP fluorescence of CcaS-CcaR system under green light, red light and darkness.
As is shown in Figure 1, in E.coli strain JT2, PcpcG2 produces 266.0 ± 19.3 and 509.0 ± 55.4 au of sfGFP in red and green light, corresponding to 1.91 ± 0.34-fold activation, demonstrating that PcpcG2 is functionally regulated by light, green-activated and red-repressed.
However, the figure also shows that that leaked expression is severe in dark surroundings. To better respond to red and green light, we optimized the promoter by refactoring it and created an innovative promoter, PcpcG2-172, with the truncation of a constitutive promoter within the cpcG2 promoter (BBa_K2012015. For more detail, please view our wiki: http://2016.igem.org/Team:HZAU-China/Experiments)
Improve by HZAU-China 2017, designed by Chu Pan
This year we improved the CcaS-CcaR system used in last year. By changing the RBS strength of CcaR, we optimized the expression of sfGFP from 2-fold variance up to 3-fold variance. For more information, please view17_HZAU-China/Improve
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]