Part:BBa_K2012015
PcpcG2-172, a modified PcpcG2 promoter
PcpcG2 promoter is a 238bp green-light activated promoter in Synechocystis PCC 6803(Part:BBa_K592003). The full length promoter is comprised of a G-box region, a CcaR-P activated promoter, and a constitutive promoter, which contributes to the leakiness under red light and low dynamic range. Therefore, we constructed PcpcG2-172, a 172bp truncated cpcG2 promoter deleted for the constitutive promoter.
Experience
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)
Fig.1 Mean sfGFP fluorescence of CcaS-CcaR system under green light, red light and darkness.
As is shown in the figure, 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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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