Difference between revisions of "Part:BBa K2384018"

 
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They have been designed for high expression in B. megaterium.The periplasm display protein Dsba-MBP, which binding the lead of the enviroment. Disulfide bond formation protein A (DsbA) is a powerful oxidative protein of the thioredoxin family. DsbA can help protein to fold correctly both in vivo and in vitro 。The C-terminal fusion protein of the maltose-binding protein can be efficiently and rapidly output into the periplasm as a key component of the expression of the metal-binding peptide of the cyclin. This year our parts were doing codon optimisation but without terminator codon.
 
They have been designed for high expression in B. megaterium.The periplasm display protein Dsba-MBP, which binding the lead of the enviroment. Disulfide bond formation protein A (DsbA) is a powerful oxidative protein of the thioredoxin family. DsbA can help protein to fold correctly both in vivo and in vitro 。The C-terminal fusion protein of the maltose-binding protein can be efficiently and rapidly output into the periplasm as a key component of the expression of the metal-binding peptide of the cyclin. This year our parts were doing codon optimisation but without terminator codon.
  
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<!-- Add more about the biology of this part here
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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--Lpp-OmpA-MBP(Pb)1.png|thumb|400px|'''Before Codon Adjustment''']]</th>
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</tr></table>
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<table><tr>
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<th>[[Image:T--FAFU-CHINA--Lpp-OmpA-MBP(Pb)2.png|thumb|400px|'''After Codon Adjustment''']]</th>
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</tr></table>
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2.Codon Used Distribution
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Show the relative codon used distribution
 +
 +
<table><tr>
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<th>[[Image:T--FAFU-CHINA--Lpp-OmpA-MBP(Pb)3.png|thumb|400px|'''Before Optimization''']]</th>
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</tr></table>
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<table><tr>
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<th>[[Image:T--FAFU-CHINA--Lpp-OmpA-MBP(Pb)4.png|thumb|400px|'''After Optimization''']]</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--Lpp-OmpA-MBP(Pb)5.png|thumb|400px|'''Before Optimization''']]</th>
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</tr></table>
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<table><tr>
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<th>[[Image:T--FAFU-CHINA--Lpp-OmpA-MBP(Pb)6.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--Lpp-OmpA-MBP SDS PAGE.png|thumb|400px|'''The expression of Lpp-OmpA-MBP 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-Lpp-OmpA+MBP(Pb).png|thumb|400px|'''Plasmid digested by PvuII''']]</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 17:54, 1 November 2017


Repressor gene xylR+Strong PxylA promoter+RBS+DsbA-MBP(Pb)+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.The periplasm display protein Dsba-MBP, which binding the lead of the enviroment. Disulfide bond formation protein A (DsbA) is a powerful oxidative protein of the thioredoxin family. DsbA can help protein to fold correctly both in vivo and in vitro 。The C-terminal fusion protein of the maltose-binding protein can be efficiently and rapidly output into the periplasm as a key component of the expression of the metal-binding peptide of the cyclin. 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.


Before Codon Adjustment
After Codon Adjustment


2.Codon Used Distribution Show the relative codon used distribution

Before Optimization
After Optimization

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

Before Optimization
After Optimization
The expression of Lpp-OmpA-MBP detected using SDS-PAGE
Plasmid digested by PvuII
Toggle switch

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 1668
    Illegal BglII site found at 1680
    Illegal XhoI site found at 847
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]