Difference between revisions of "Part:BBa K3712013"

 
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<partinfo>BBa_K3712013 short</partinfo>
 
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===USAGE AND RESULTS===
  
 
PR/PL, a tandem PR and PL promoter from lambda phage, to ensure high-level expression of genes.Our target gene was cloned downstream of the pL and/or pR promoters. When cultured at 37&#8451;, the expression of target gene is inhibited by Tcl and the target protein is hardly expressed. Under the induction of temperature at 45&#8451; or infrared light which increases temperature above 37&#8451;, Tcl degrades and the inhibition of the promoter releases. So, the expression of target protein is initiated
 
PR/PL, a tandem PR and PL promoter from lambda phage, to ensure high-level expression of genes.Our target gene was cloned downstream of the pL and/or pR promoters. When cultured at 37&#8451;, the expression of target gene is inhibited by Tcl and the target protein is hardly expressed. Under the induction of temperature at 45&#8451; or infrared light which increases temperature above 37&#8451;, Tcl degrades and the inhibition of the promoter releases. So, the expression of target protein is initiated
  
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===RESULTS===
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===Usage and Biology===
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<title>Figure 1. The agarose gel electrophoresis map of Wild-type BLP-7-27 ELP, Optimized BLP-7-27 ELP, Wild-type TLR2-27ELP, and Optimized TLR2-27 ELP respectively constructed in pR/pL expression system. </title>
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<center><img src="https://2021.igem.org/wiki/images/4/45/T--HUST2-China--The_agarose_gel_electrophoresis_map.png" style="width:793px;height:360px"></center>
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<center><b>Figure 2.</b> About 200-1000 ng of plasmid was digested at 37℃ for 30-60 minutes and analyzed on 1% Agarose Gel. Lane1: circular plasmid (brighter) and supercoiled structure; Lane2: plasmid (brighter) digested by EcoRI and BamHI and target gene fragment. </center>
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Revision as of 07:44, 21 October 2021


pR/pL promotor

USAGE AND RESULTS

PR/PL, a tandem PR and PL promoter from lambda phage, to ensure high-level expression of genes.Our target gene was cloned downstream of the pL and/or pR promoters. When cultured at 37℃, the expression of target gene is inhibited by Tcl and the target protein is hardly expressed. Under the induction of temperature at 45℃ or infrared light which increases temperature above 37℃, Tcl degrades and the inhibition of the promoter releases. So, the expression of target protein is initiated

RESULTS

Figure 1. The agarose gel electrophoresis map of Wild-type BLP-7-27 ELP, Optimized BLP-7-27 ELP, Wild-type TLR2-27ELP, and Optimized TLR2-27 ELP respectively constructed in pR/pL expression system.

Figure 2. About 200-1000 ng of plasmid was digested at 37℃ for 30-60 minutes and analyzed on 1% Agarose Gel. Lane1: circular plasmid (brighter) and supercoiled structure; Lane2: plasmid (brighter) digested by EcoRI and BamHI and target gene fragment.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 116
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]