Difference between revisions of "Part:BBa K2933215"
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'''Figure 1.'''    The PCR result of BcII.<br>
 
'''Figure 1.'''    The PCR result of BcII.<br>
  
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Then we used the vector pGEX-6p-1 to construct our expression plasmid. And then we converted the plasmid constructed to ''E. coli'' DH5α to expand the plasmid largely.After verification, it was determined that the construction is successful. We converted the plasmid to ''E. coli'' BL21(DE3) for expression and purification.<br>
 
  
  
 
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Revision as of 13:03, 23 September 2019


T7 promoter+RBS b+Linker h+His+Linker f+BCll-194+T7 terminator

This part consists of T7 promoter, RBS and protein coding sequence（His+Linker f+NDM-23），and the biological module can be built into E.coli for protein expression.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 47
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 169
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 47
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 47
  • 1000
    COMPATIBLE WITH RFC[1000]


Usage and Biology=

This composite part is made up with seven basic parts, the His tag, T7 promoter, RBS and our target protein BcII-194. It encodes a protein which is BcII-194 fused with His tag. The fusion protein is about 28.1 kD. In order to gain the highly purified target protein, we add His tag in N-terminal of BcII-194. It is convenient for us to purify our target protein.

Molecular cloning

First,we obtained BcII by PCR.

BcII-194 PCR.jpeg
Figure 1. The PCR result of BcII.