Part:BBa_K2933160
His+Linker f+Fla.103
This part encodes the fusion protein of His tag and Fla.103 to promote the expression and purification of target protein(Fla.103).
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 162
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 51
Illegal PstI site found at 162 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 644
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 162
- 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 162
- 1000COMPATIBLE WITH RFC[1000]
Usage and Biology
This composite part is made up with three basic parts, the HiS tag, the thrombin restriction site and our target protein Fla.103. It encodes a protein which is Fla.103 fused with His tag. The fusion protein is about 28.7 kD. It is convenient for us to purify our target protein.
Molecular cloning
First, 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.We then extracted plasmids and performed double enzyme digestion verification.
Figure 1. (a) The PCR result of Fla.103. (b) The verification results by enzyme digestion.The 1 is the original plasmid.The 2 is the
results verified by double 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.
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