Difference between revisions of "Part:BBa K2721010:Design"

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===Design Notes===
 
===Design Notes===
−
We obtained this part by PCR amplification and verified it by agarose gel electrophoresis. Then we assembled it into pSB1C3 and expressed the csgA in DH5alpha.We used the cnago red to dye the pellet and proved the expression.(Result is as follow)
+
We obtained this part by PCR amplification and added spytag in the N-terminal and 6Xhistag in C-terminal in the PCR primers.  After verifying it by agarose gel electrophoresis，we assembled it into pSB1C3 and expressed the csgA in DH5alpha.We used the cnago red to dye the pellet and proved the expression.(Result is as follow)
  
 
===Source===
 
===Source===
  
−
We obtained this part by PCR amplification from the genome of Escherichia coli MG1655.
+
We obtained this part by PCR amplification from the genome of Escherichia coli MG1655.They we added the spytag in the N-terminal and 6Xhistag in C-terminal in the PCR primers.
  
 
===References===
 
===References===
 
[1] Edwin van Bloois, Remko T. Winter, et al. Decorating microbes: surface display of proteins on Escherichia coli.Cell, 2011,29(2):79-86
 
[1] Edwin van Bloois, Remko T. Winter, et al. Decorating microbes: surface display of proteins on Escherichia coli.Cell, 2011,29(2):79-86
 
[2] Michelle M. Barnhart, Matthew R. Chapman. Curli Biogenesis and Function. Annu. Rev. Microbiol. 2006. 60:131–47
 
[2] Michelle M. Barnhart, Matthew R. Chapman. Curli Biogenesis and Function. Annu. Rev. Microbiol. 2006. 60:131–47

Latest revision as of 08:01, 4 October 2018


the gene of csgA


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

We obtained this part by PCR amplification and added spytag in the N-terminal and 6Xhistag in C-terminal in the PCR primers. After verifying it by agarose gel electrophoresis，we assembled it into pSB1C3 and expressed the csgA in DH5alpha.We used the cnago red to dye the pellet and proved the expression.(Result is as follow)

Source

We obtained this part by PCR amplification from the genome of Escherichia coli MG1655.They we added the spytag in the N-terminal and 6Xhistag in C-terminal in the PCR primers.

References

[1] Edwin van Bloois, Remko T. Winter, et al. Decorating microbes: surface display of proteins on Escherichia coli.Cell, 2011,29(2):79-86 [2] Michelle M. Barnhart, Matthew R. Chapman. Curli Biogenesis and Function. Annu. Rev. Microbiol. 2006. 60:131–47