Difference between revisions of "Part:BBa K2012015"

 
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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.
 
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.
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<p class="MsoNormal">Experience</p>
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<p class="MsoNormal">&nbsp;</p>
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<p class="MsoNormal">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)</p>
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<p class="MsoNormal">&nbsp;</p>
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<p class="MsoNormal"><img src="https://static.igem.org/mediawiki/parts/d/db/Hzau-china_cpcg2.jpg" alt="00.1" width="480" height="293" /></p>
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<p class="MsoNormal"><strong>Fig.1 <a name="OLE_LINK16" id="OLE_LINK16"></a><a name="OLE_LINK15" id="OLE_LINK15">Mean  sfGFP fluorescence of CcaS-CcaR</a> system under green light, red light  and darkness.</strong></p>
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<p class="MsoNormal">&nbsp;</p>
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<p class="MsoNormal">As is shown in the figure, in E.coli strain  JT2, PcpcG2 produces 266.0 &plusmn; 19.3 and 509.0 &plusmn; 55.4 au of sfGFP in red and green  light, corresponding to 1.91 &plusmn; 0.34-fold activation, demonstrating that PcpcG2  is functionally regulated by light, green-activated and red-repressed.</p>
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<p class="MsoNormal">&nbsp;</p>
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<p class="MsoNormal">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). </p>
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===Usage and Biology===
 
===Usage and Biology===

Revision as of 10:26, 14 October 2016


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)

 

00.1

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


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]