Part:BBa_K2933145
His+Linker f+MYX-1
This part encodes the fusion protein of His tag and MYX-1 to promote the expression and purification of target protein(MYX-1).
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 166
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 51
Illegal PstI site found at 166 - 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 166
- 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 166
- 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 MYX-1. It encodes a protein which is MYX-1 fused with His tag. The fusion protein is about 27.9 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.
Figure 1. Left: The PCR result of MYX-1. 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
[1]Correction for Goldman et al., Evolution of sensory complexity recorded in a myxobacterial genome. Proc Natl Acad Sci U S A. 2006;103(51):19605. doi:10.1073/pnas.0609567103.
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