Difference between revisions of "Part:BBa K2012015"
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<h3><b>Author: Kening Chen</b></h3>
 
<h3><b>Author: Kening Chen</b></h3>
 
<p>&nbsp;&nbsp;&nbsp;&nbsp;Measurement of PcpcG2-172 under new circumstances.</p>
 
<p>&nbsp;&nbsp;&nbsp;&nbsp;Measurement of PcpcG2-172 under new circumstances.</p>
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<p>&nbsp;&nbsp;&nbsp;&nbsp; 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>
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<p>&nbsp;&nbsp;&nbsp;&nbsp; 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>

Revision as of 05:16, 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


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]

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.