Difference between revisions of "Part:BBa K4054100"
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pLuxR is a common used induciable promoter. However, almost all of the teams characterized the pLuxR basing on AHL concentration. In our project this year, depending our <i>in vitro</i> system, we measure the promoter strenth changes at different concentration of LuxR.We had expressed LuxR by BL21(DE) and harvest it, which we used in the measurement by diluting LuxR to different concentrations and added into the cell free system. Through real-time detection by microplate reader, we achieved the result and confirmed the transition concerntration range.
 
pLuxR is a common used induciable promoter. However, almost all of the teams characterized the pLuxR basing on AHL concentration. In our project this year, depending our <i>in vitro</i> system, we measure the promoter strenth changes at different concentration of LuxR.We had expressed LuxR by BL21(DE) and harvest it, which we used in the measurement by diluting LuxR to different concentrations and added into the cell free system. Through real-time detection by microplate reader, we achieved the result and confirmed the transition concerntration range.
  
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[[File:T--XJTLU-CHINA-RT.png|400px|thumb|left|alt=domestication.|Experimental results for characterization of pLux promoter.]]
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[[File:T--XJTLU-CHINA-RT.png|600px|thumb|center|alt=domestication.|Experimental results for characterization of pLux promoter.]]
  
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[[File:T--XJTLU-CHINA-S Curve.png|400px|thumb|alt=domestication.|Final GFP expression levels for different AHL inducer concentrations.]]
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[[File:T--XJTLU-CHINA-S Curve.png|600px|thumb|center|alt=domestication.|Final GFP expression levels for different AHL inducer concentrations.]]
 
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Revision as of 20:12, 19 October 2021


pLuxR_T7 RBS_GFP

demostication.
Figure 1. The promoter pLuxR could be activated by AHL and then start the expression of eGFP.

This is a composite part used for GFP expression. It consist of pLuxR, T7 RBS and ther coding sequence of reporter GFP As it include the component "pLuxR", the activator LuxR is required to start the transcription.

Introduction

In this year, XJTLU-CHINA characterized pLuxR promoter by this part. As the Figure 1 shows, the composite part consists of the promoter pLuxR (K4054017), the RBS (K4054017)，the coding sequence of eGFP (E0040) and the terminator K4054018.


The part was expressed in Cell Free systemm (in vitro transcription and translation system)， which allowed us to measure the GFP expression level under different LuxR concentration


Experiment

pLuxR is a common used induciable promoter. However, almost all of the teams characterized the pLuxR basing on AHL concentration. In our project this year, depending our in vitro system, we measure the promoter strenth changes at different concentration of LuxR.We had expressed LuxR by BL21(DE) and harvest it, which we used in the measurement by diluting LuxR to different concentrations and added into the cell free system. Through real-time detection by microplate reader, we achieved the result and confirmed the transition concerntration range.

domestication.
Experimental results for characterization of pLux promoter.
domestication.
Final GFP expression levels for different AHL inducer concentrations.


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
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 736