Difference between revisions of "Part:BBa K2384013"
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<p>Xylose-inducible expression system: strong<b> P<i>xylA</i></b> promoter and repressor gene <b><i>xylR</i></b>; after xylose addition, the repressor is released from the P<i>xylA</i> that activates transcription initiation.([[Part:BBa_K733002|BBa_K733002]]) They have been designed for high expression in <i>B. megaterium</i> This part serves as a functional MT for heavy metal binding and conferred resistance toxic metal cadmium. GST as a fusion partenr is to enhance stability of expression.The original part was from Cornell([[Part:BBa_K1460002|BBa_K1460002]]). This year our parts were doing codon optimisation but without terminator codon.</p>
 
<p>Xylose-inducible expression system: strong<b> P<i>xylA</i></b> promoter and repressor gene <b><i>xylR</i></b>; after xylose addition, the repressor is released from the P<i>xylA</i> that activates transcription initiation.([[Part:BBa_K733002|BBa_K733002]]) They have been designed for high expression in <i>B. megaterium</i> This part serves as a functional MT for heavy metal binding and conferred resistance toxic metal cadmium. GST as a fusion partenr is to enhance stability of expression.The original part was from Cornell([[Part:BBa_K1460002|BBa_K1460002]]). This year our parts were doing codon optimisation but without terminator codon.</p>
  
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===Usage and Biology===
 
===Usage and Biology===
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<b>Optimization Report</b>
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1.Codon Used Adjustment
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The best value is 1 for sequence optimization.
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<table><tr>
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<th>[[Image:T--FAFU-CHINA--GST-CRS51.png|thumb|400px|'''After Codon Adjustment''']]</th>
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</tr></table>
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3.GC Content:
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The comparison of  GC content between original sequence and optimized sequence
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 +
<table><tr>
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<th>[[Image:T--FAFU-CHINA--GST-CRS52.png|thumb|400px|'''After Optimization''']]</th>
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</tr></table>
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<table><tr>
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<th>[[Image:T--FAFU-CHINA--GST-CRS5 SDS PAGE.jpg|thumb|400px|'''The expression of GST-CRS5 detected using SDS-PAGE ''']]</th>
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</tr></table>
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<table><tr>
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<th>[[Image:T--FAFU-CHINA--M-GST-CRS5.jpg|thumb|400px|'''Plasmid digested by EcoRI and HindIII ''']]</th>
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</tr></table>
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<table><tr>
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<th>[[Image:T--FAFU-CHINA--GST-CRS5-Toggle-switch.png|thumb|400px|'''Toggle switch ''']]</th>
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</tr></table>
  
 
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Revision as of 17:49, 1 November 2017


Repressor gene xylR+Strong PxylA promoter+RBS+GST-CRS5+His-tag

Xylose-inducible expression system: strong PxylA promoter and repressor gene xylR; after xylose addition, the repressor is released from the PxylA that activates transcription initiation.(BBa_K733002) They have been designed for high expression in B. megaterium This part serves as a functional MT for heavy metal binding and conferred resistance toxic metal cadmium. GST as a fusion partenr is to enhance stability of expression.The original part was from Cornell(BBa_K1460002). This year our parts were doing codon optimisation but without terminator codon.


Usage and Biology

Optimization Report

1.Codon Used Adjustment The best value is 1 for sequence optimization.

After Codon Adjustment


3.GC Content: The comparison of GC content between original sequence and optimized sequence

After Optimization
The expression of GST-CRS5 detected using SDS-PAGE
Plasmid digested by EcoRI and HindIII
Toggle switch

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 2159
    Illegal XhoI site found at 847
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 1571