Difference between revisions of "Part:BBa K2012015"
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<p> PcpcG2-172 is a 172bp green-light induced promoter constructed by Sebastian R. Schmidl group (1)[https://parts.igem.org/(1)].CcaS phosphorylates CcaR under green light and induces expression of cpcG2 (2)[https://parts.igem.org/(2)].A study of Mitsuharu Nakajima group reveals that CcaS#10 works well when two PAS domains were deleted from the original sequence (3)[https://parts.igem.org/(3)].Therefore, we tested PcpcG2-172 in a modified two-component system composed of CcaS#10 and a series of CcaRs with different transcription intensities.</p> | <p> PcpcG2-172 is a 172bp green-light induced promoter constructed by Sebastian R. Schmidl group (1)[https://parts.igem.org/(1)].CcaS phosphorylates CcaR under green light and induces expression of cpcG2 (2)[https://parts.igem.org/(2)].A study of Mitsuharu Nakajima group reveals that CcaS#10 works well when two PAS domains were deleted from the original sequence (3)[https://parts.igem.org/(3)].Therefore, we tested PcpcG2-172 in a modified two-component system composed of CcaS#10 and a series of CcaRs with different transcription intensities.</p> | ||
| − | [[File:T—HZAU-China—Figure1.png]] | + | [[File:T—HZAU-China—Figure1.png|600px|thumb|left|Figure1. Light inducible CcaS-CcaR two component system. (A) Structure of the CcaS-CcaR circuit. (B) Measurement of PcpcG2-172 under red light (660nm) or green light (520nm). The experiment was carried out in MG1655-△EnvZ strain, using a 24-wall plate adapted LED device. Graphic shows that among four tested promoters, J23117-CcaR gives PcpcG2-172 best dynamic range of 8.3-fold change. When using stronger promoters J23114 and J23115, this dynamic range narrows as leakage under red light getting larger.]] |
Revision as of 08:43, 1 October 2021
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.(For more detail, please view our wiki: "http://2016.igem.org/Team:HZAU-China/Experiments" )
Figure 1. Fluorescence assay of CcaS-CcaR system with PcpcG2-172 (BBa_K2012015) in CL1 (△EnvZ), and PCB (△CcaS) as chromophore.
Fluorescence under green light is 1.81-folds of red light, proving that green light activates output expression, the device works well. PcpcG2-172 shows high efficiency.
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]
iGEM 2021 XHD-Wuhan-B-China, new documentation (For Bronze)
Group: XHD-Wuhan-B-China
Author: Kening Chen
Measurement of PcpcG2-172 under new circumstances.
PcpcG2-172 is a 172bp green-light induced promoter constructed by Sebastian R. Schmidl group (1)[1].CcaS phosphorylates CcaR under green light and induces expression of cpcG2 (2)[2].A study of Mitsuharu Nakajima group reveals that CcaS#10 works well when two PAS domains were deleted from the original sequence (3)[3].Therefore, we tested PcpcG2-172 in a modified two-component system composed of CcaS#10 and a series of CcaRs with different transcription intensities.
