Difference between revisions of "Part:BBa K2384018"
 
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<b>Optimization Report</b>
 
<b>Optimization Report</b>
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Gene Name:DSBA-MBP(Pb)
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Expression System:Bacillus megaterium
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Gene Length:963 (bp)
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1.Codon Used Adjustment
 
1.Codon Used Adjustment
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<table><tr>
 
<table><tr>
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<th>[[Image:T--FAFU-CHINA--DSBA-MBP(Pb)1.png|thumb|400px|'''Before Codon Adjustment''']]</th>
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<th>[[Image:T--FAFU-CHINA--DSBA-MBP(Pb)1.png|thumb|400px|'''Figure 1:Before Codon Adjustment''']]</th>
 
</tr></table>
 
</tr></table>
  
 
<table><tr>
 
<table><tr>
−
<th>[[Image:T--FAFU-CHINA--DSBA-MBP(Pb)2.png|thumb|400px|'''After Codon Adjustment''']]</th>
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<th>[[Image:T--FAFU-CHINA--DSBA-MBP(Pb)2.png|thumb|400px|'''Figure 2:After Codon Adjustment''']]</th>
 
</tr></table>
 
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<table><tr>
 
<table><tr>
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<th>[[Image:T--FAFU-CHINA--DSBA-MBP(Pb)3.png|thumb|400px|'''Before Optimization''']]</th>
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<th>[[Image:T--FAFU-CHINA--DSBA-MBP(Pb)3.png|thumb|400px|'''Figure 3:Before Optimization''']]</th>
 
</tr></table>
 
</tr></table>
  
 
<table><tr>
 
<table><tr>
−
<th>[[Image:T--FAFU-CHINA--DSBA-MBP(Pb)4.png|thumb|400px|'''After Optimization''']]</th>
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<th>[[Image:T--FAFU-CHINA--DSBA-MBP(Pb)4.png|thumb|400px|'''Figure 4:After Optimization''']]</th>
 
</tr></table>
 
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<table><tr>
 
<table><tr>
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<th>[[Image:T--FAFU-CHINA--DSBA-MBP(Pb)5.png|thumb|400px|'''Before Optimization''']]</th>
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<th>[[Image:T--FAFU-CHINA--DSBA-MBP(Pb)5.png|thumb|400px|'''Figure 5:Before Optimization''']]</th>
 
</tr></table>
 
</tr></table>
  
 
<table><tr>
 
<table><tr>
−
<th>[[Image:T--FAFU-CHINA--DSBA-MBP(Pb)(Pb)6.png|thumb|400px|'''After Optimization''']]</th>
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<th>[[Image:T--FAFU-CHINA--DSBA-MBP(Pb)(Pb)6.png|thumb|400px|'''Figure 6:After Optimization''']]</th>
 
</tr></table>
 
</tr></table>
  
 
<table><tr>
 
<table><tr>
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<th>[[Image:T--FAFU-CHINA--DsbA-MBP123.jpg|thumb|400px|'''The expression of DsbA+MBP(Pb) detected using SDS-PAGE ''']]</th>
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<th>[[Image:T--FAFU-CHINA--DsbA-MBP123.jpg|thumb|400px|'''Figure 7:The expression of DsbA+MBP(Pb) detected using SDS-PAGE ''']]</th>
 
</tr></table>
 
</tr></table>
  
 
<table><tr>
 
<table><tr>
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<th>[[Image:T--FAFU-CHINA--M-DSBA-MBP(Pb).png|thumb|400px|'''Plasmid digested by EcoRI-PstI ''']]</th>
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<th>[[Image:T--FAFU-CHINA--M-DSBA-MBP(Pb).png|thumb|400px|'''Figure 8:Plasmid digested by EcoRI-PstI ''']]</th>
 
</tr></table>
 
</tr></table>
  
 
<table><tr>
 
<table><tr>
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<th>[[Image:T--FAFU-CHINA--GST-CRS5-Toggle-switch.png|thumb|400px|'''Our genetic circuit ''']]</th>
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<th>[[Image:T--FAFU-CHINA--GST-CRS5-Toggle-switch.png|thumb|400px|'''Figure 9:Our genetic circuit ''']]</th>
 
</tr></table>
 
</tr></table>
  

Latest revision as of 19:08, 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

Gene Name:DSBA-MBP(Pb)

Expression System:Bacillus megaterium

Gene Length:963 (bp)


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


Figure 1:Before Codon Adjustment
Figure 2:After Codon Adjustment


2.Codon Used Distribution Show the relative codon used distribution

Figure 3:Before Optimization
Figure 4:After Optimization

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

Figure 5:Before Optimization
Figure 6:After Optimization
Figure 7:The expression of DsbA+MBP(Pb) detected using SDS-PAGE
Figure 8:Plasmid digested by EcoRI-PstI
Figure 9:Our genetic circuit

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]