Part:BBa_K2933159
His+Linker f+IMP-71
This part encodes the fusion protein of His tag and IMP-71 to promote the expression and purification of target protein(IMP-71).
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 810
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 51
Illegal PstI site found at 810 - 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 810
- 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 810
- 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 IMP-71. It encodes a protein which is IMP-71 fused with His tag. The fusion protein is about 27.2 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. The PCR result of IMP-71.
References
[1]Purification, crystallization and preliminary X-ray analysis of IMP-18, a class B carbapenemase from Pseudomonas aeruginosa.Furuyama T, Ishii Y, Ohya N, Tateda K, Hanson ND, Shimizu-Ibuka A.Acta Crystallogr Sect F Struct Biol Cryst Commun. 2013 Dec;69(Pt 12):1397-400.
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