Difference between revisions of "Part:BBa K2384004"
 
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DsbA, a protein that can export its C-terminal fusion protein into periplasm more efficiently and faster than maltose binding protein, was used as the key component to express our Metal Binding Peptide periplasmically .
 
DsbA, a protein that can export its C-terminal fusion protein into periplasm more efficiently and faster than maltose binding protein, was used as the key component to express our Metal Binding Peptide periplasmically .
  
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Our sequence optimization is based on the sequence provided by Peking University's IGEM VIKI in 2010.It is more suitable for expression in Bacillus  
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Our sequence optimization is based on the sequence provided by Peking University's IGEM Wiki in 2010. ([[Part:BBa_K346003|BBa_K346003]])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===
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<b>Optimization Report</b>
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Gene Name:DSBA-MBP(Hg)
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Expression System:Bacillus megaterium
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Gene Length:963 (bp)
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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--DSBA-MBP(Hg)1.png|thumb|400px|'''Figure 1: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--DSBA-MBP(Hg)2.png|thumb|400px|'''Figure 2: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
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<table><tr>
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<th>[[Image:T--FAFU-CHINA--DSBA-MBP(Hg)3.png|thumb|400px|'''Figure 3:Before Optimization''']]</th>
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</tr></table>
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<table><tr>
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<th>[[Image:T--FAFU-CHINA--DSBA-MBP(Hg)4.png|thumb|400px|'''Figure 4: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--DSBA-MBP(Hg)5.png|thumb|400px|'''Figure 5:Before Optimization''']]</th>
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</tr></table>
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<table><tr>
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<th>[[Image:T--FAFU-CHINA--DSBA-MBP(Hg)6.png|thumb|400px|'''Figure 6:After Optimization''']]</th>
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</tr></table>
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<table><tr>
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<th>[[Image:T--FAFU-CHINA--DsbA-MBP123.jpg|thumb|400px|'''Figure 7:The expression of DsbA-MBP(Hg) 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-DSBA-MBP(Hg).png|thumb|400px|'''Figure 8:Plasmid digested by PvuII ''']]</th>
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</tr></table>
  
 
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Latest revision as of 22:18, 1 November 2017


DsbA+MBP(Hg)

DsbA, a protein that can export its C-terminal fusion protein into periplasm more efficiently and faster than maltose binding protein, was used as the key component to express our Metal Binding Peptide periplasmically .

Our sequence optimization is based on the sequence provided by Peking University's IGEM Wiki in 2010. (BBa_K346003)The optimization of this sequence is more suitable for the expression of Bacillus megaterium.



Usage and Biology

Optimization Report

Gene Name:DSBA-MBP(Hg)

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(Hg) detected using SDS-PAGE
Figure 8:Plasmid digested by PvuII

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
    Illegal BglII site found at 711
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]