Difference between revisions of "Part:BBa K2933265"

Latest revision as of 14:07, 24 September 2019

RBS+Linker h+His+Linker f+NDM-23+T7 terminator

This part consists of RBS, Linker h, protein coding sequence(His+Linker f+NDM-23) and T7 terminator,and the biological module can be build into E.coli for protein expression. This part can be prefaced with promoters of different strengths and types to regulate expression function.

Sequence and Features

Usage and Biology

This composite part is made up with six basic parts(RBS , Linker h, His, Linker f , NDM-23 and T7 terminator ). It encodes a protein which is NDM-23 fused with His tag. The fusion protein is about 28.5 kD. It is convenient for us to purify our target protein.

Molecular cloning

We used the vector pET28a to construct our expression plasmid. And then we converted the plasmid constructed to E. coli DH5α to expand the plasmid largely.

Figure 1. Left: The PCR result of NDM-23. Right: The verification results by enzyme digestion.

After verification, it was determined that the construction is successful. We converted the plasmid to E. coli BL21(DE3) for expression and purification.

References

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