Difference between revisions of "Part:BBa K2012015"
 
(50 intermediate revisions by 6 users not shown)
Line 2: Line 2:
 
__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K2012015 short</partinfo>
 
<partinfo>BBa_K2012015 short</partinfo>
 +
<p>
 +
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" )
 +
</p>
 +
<html>
  
−
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.
+
<div>
−
<html xmlns="http://www.w3.org/1999/xhtml">
+
<div style="height:auto!important;margin-left:auto;margin-right:auto;min-height:841.9pt;padding-bottom:1pt;padding-top:1pt">
−
<head>
+
<div style="height:auto!important;margin-left:auto;margin-right:auto;min-height:841.9pt;padding-bottom:1pt;padding-top:1pt">     
−
<meta http-equiv="Content-Type" content="text/html; charset=utf-8" />
+
<div style="margin-left:auto;margin-right:auto;position:relative">
 +
<div style="height:auto!important;margin-left:auto;margin-right:auto;min-height:841.9pt;padding-bottom:1pt;padding-top:1pt">     
 +
<div style="margin-left:auto;margin-right:auto;position:relative">
 +
<div style="height:auto!important;margin-left:auto;margin-right:auto;min-height:841.95pt;padding-bottom:1pt;padding-top:1pt">     
 +
<div style="margin-left:auto;margin-right:auto;position:relative">
 +
<div style="height:auto!important;margin-left:auto;margin-right:auto;min-height:841.9pt;padding-bottom:1pt;padding-top:1pt">     
 +
<div style="margin-left:auto;margin-right:auto;position:relative"><p style="text-align:justify;text-justify:inter-ideograph"><span style="font-family:Calibri"><img height="318.92999267578125" src="https://static.igem.org/mediawiki/2016/5/5f/Eeeee.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">&nbsp;</span></p>
  
−
<style type="text/css">
+
<p style="text-align:justify;text-justify:inter-ideograph"><span style="font-family:Calibri"><strong>Figure 1.</strong> Fluorescence assay of CcaS-CcaR system with PcpcG2-172 (BBa_K2012015) in CL1 (△EnvZ), and PCB (△CcaS) as </span><span style="font-family:Calibri">chromophore</span><span style="font-family:Calibri">. </span><span style="font-family:Calibri"><span></span></span></p>
−
<!--
+
<p style="text-align:justify;text-justify:inter-ideograph"><span style="font-family:Calibri">F</span><span style="font-family:Calibri">luorescence under green light is 1.81-folds of red light, proving that green light activates output expression, the device works well. </span><span style="font-family:Calibri">PcpcG2-172 </span><span style="font-family:Calibri">shows high efficiency.<span></span></span></p>
−
p.MsoNormal {
+
</br>
−
margin:0cm;
+
</br>
−
margin-bottom:.0001pt;
+
</br>
−
text-align:justify;
+
−
text-justify:inter-ideograph;
+
−
font-size:10.5pt;
+
−
font-family:"Calibri",sans-serif;
+
−
}
+
−
-->
+
−
</style>
+
−
</head>
+
−
 
+
−
<body>
+
−
<p class="MsoNormal">Experience</p>
+
−
<p class="MsoNormal">&nbsp;</p>
+
−
<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>
+
−
<p class="MsoNormal">&nbsp;</p>
+
−
<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>
+
−
<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>
+
−
<p class="MsoNormal">&nbsp;</p>
+
−
<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>
+
−
<p class="MsoNormal">&nbsp;</p>
+
−
<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>
+
−
</body>
+
 
</html>
 
</html>
−
<!-- Add more about the biology of this part here
 
−
===Usage and Biology===
 
  
 +
<b>Improvement</b><br>A truncated promoter PcpcG2-68 has the similar pattern under the control of green/red light as PcpcG2-172, but is shorter and more convenient to construct. For example, when constructing an array of sgRNAs, PcpcG2-68 has a big advantage over PcpcG2-172: reducing the volume of the circuit and cheaper to synthesis.<br> [[File:T--XHD-Wuhan-B-China--111(1).png|900px|thumb|left|]]<br>Please view https://parts.igem.org/Part:BBa_K3921000 for more details.
 
<!-- -->
 
<!-- -->
 
<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
Line 47: Line 36:
 
<partinfo>BBa_K2012015 parameters</partinfo>
 
<partinfo>BBa_K2012015 parameters</partinfo>
 
<!-- -->
 
<!-- -->
 +
 +
<span id="IGEM 2021 XHD- Wuhan-B-China"><br><br><br><br><br></span>

Latest revision as of 06:41, 21 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.




Improvement
A truncated promoter PcpcG2-68 has the similar pattern under the control of green/red light as PcpcG2-172, but is shorter and more convenient to construct. For example, when constructing an array of sgRNAs, PcpcG2-68 has a big advantage over PcpcG2-172: reducing the volume of the circuit and cheaper to synthesis.
T--XHD-Wuhan-B-China--111(1).png

Please view https://parts.igem.org/Part:BBa_K3921000 for more details.

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]