Difference between revisions of "Part:BBa K2012023"

Revision as of 14:53, 1 November 2016

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)

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

@@ Line 13: / Line 13: @@
 <p style="text-align:justify;text-justify:inter-ideograph"><span style="font-family:Calibri">&nbsp;</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"><b>Figure 1.</b> Mean 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"><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">&nbsp;</span></p>
 <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>