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| − | <b>RpaR-pRpa-GFP</b><br> | + | <b>Team:Shanghaitech</b> |
| − | Group: <b>Shanghaitech iGEM 2017</b>
| + | <b>RpaR-RHOPA-pLas-GFP</b> |
| | + | Quorum sensing receivers and non-cognate promotor test bed for receiver protein -- promotor orthogonality testing. Consist of a Quorum sensing receiver CDS and a GFP CDS controlled by non-cognate Quorum sensing promotors. |
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| − | The HSL receiver RpaR from P.aeruginosa activates expression of GFP protein in response to Coumaroyl-HSL.
| + | [[File:T--Shanghaitech--QSRLF.jpeg|thumb|center|700px|<b>Fig. 1 Genetic Circuit Design</b>]] |
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| − | This Receiver can be easily replaced by other AHL receivers in our collection. A full collection could be found in: http://2017.igem.org/Team:Shanghaitech/Library
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| − | ==<b>Usage and Biology</b>==
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| − | [[File:RpaRpRpaGFPPARTS.png|thumb|center|700px|<b>Fig. 1 Genetic Circuit Design</b>]]
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| − | When AHL is added with concentration higher than a critical value, the constitutively expressed RpaR will bind to the AHL molecule Coumaroyl-HSL, dimerize and bind to the pRpa regulatory sequence to activate GFP expression.
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| − | ==<b>Fluorescent Response to cognate Coumaroyl-HSL</b>==
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| − | To test this part, we used standard Coumaroyl-HSL (HSL produced by RpaI in P.aeruginosa) to determine the response curve.
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| − | [[File:T--Shanghaitech--RpaRGFPLF.png|thumb|center|800px|<b>Fig. 1 RpaR-pRpa-GFP‘s response to cognate CoumaroylHSL </b>]] | + | |
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| − | ==Orthogonality test against non-cognate inducers==
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| − | We have characterized crosstalk response of RpaR to several non-cognate AHLs:
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| − | [[File:T--Shanghaitech--RpaRorthogonality.png|thumb|center|800px|<b>Fig. 2 Orthogonality test of RpaR-pRpa-GFP </b> (i):Fluorescent response to cognate and non-cognate AHLs (ii)Dose-Response curves for cognate and non-cognate AHLs (iii-vi)Fluorescent response to non-cognate AHLs in compared with Coumaroyl-HSL ]]
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| − | It has shown that RpaR is sensitive to it's cognate HSL and has obvious crosstalk with 3OC8-HSL in Tra System in relatively high concentration.
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| − | __NOTOC__
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| − | <partinfo>BBa_K2315006 short</partinfo>
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| − | RpaR-RHOPA-pLas-GFP
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| − | <!-- Add more about the biology of this part here
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| − | ===Usage and Biology===
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| − | <span class='h3bb'>Sequence and Features</span>
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| − | <partinfo>BBa_K2315006 SequenceAndFeatures</partinfo>
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| − | <!-- Uncomment this to enable Functional Parameter display
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| − | ===Functional Parameters===
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| − | <partinfo>BBa_K2315006 parameters</partinfo>
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| − | <!-- -->
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