Construction of prokaryotic expression plasmid of fusion protein including porin A and porin B ofNeisseria gonorrhoeae and its expression inE. coli

Abstract

In order to provide a rational research basis for clinical detection and genetic engineering vaccine, plasmid pET-28a (+) encoding both Porin gene PIA and PIB ofNeisseria gonorrhoeae was constructed and a fusion protein inE. coli DE3 expressed. The fragments of PIA and PIB gene ofNeisseria gonorrhoeae were amplified and cloned into prokaryotic expression plasmid pET-28a (+) with double restriction endonuclease cut to construct recombinant pET-PIB-PIA. The recombinant was verified with restriction endonuclease and sequenced and transformed intoE. coli DE3 to express the fusion protein PIB-PIA after induced with IPTG. The results showed PIA-PIB fusion DNA fragment was proved correct through sequencing. A 67 kD (1 kD=0.992 1 ku) fusion protein had been detected by SDS-PAGE. It was concluded that the fusion protein was successively expressed.

Keywords

Neisseria gonorrhoeae / porin B / porin A / prokaryotic expression plasmid recombinant / fusion protein

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