Difference between revisions of "Part:BBa K2384003"
 
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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.
 
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.
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Our sequence optimization is based on the sequence provided by Peking University's IGEM Wiki in 2010.It is more suitable for expression in Bacillus  
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Our sequence optimization is based on the sequence provided of Peking University's IGEM in 2010.([[Part::BBa_K346004|:BBa_K346004]])The optimization of this sequence is more suitable for the expression of <i>Bacillus megaterium</i>.
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===Usage and Biology===
 
===Usage and Biology===
  
 
<b>Optimization Report</b>
 
<b>Optimization Report</b>
 +
 +
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>
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<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>
 
</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>
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<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>
 
</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>
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<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>
  

Latest revision as of 22:08, 1 November 2017


DsbA+MBP(Pb)

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.

Our sequence optimization is based on the sequence provided of Peking University's IGEM in 2010.(:BBa_K346004)The optimization of this sequence is more suitable for the expression of Bacillus megaterium.

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


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 255
    Illegal BglII site found at 267
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]