Cloning, Expression Of Salmonella Invasion Protein (InvA) Gene And The Primary Study On The Expressed Protein

Objective:To expression InvA fusion protein in E.coli in high efficiency, the invA gene of salmonella was cloned into an expression plasmid to generate pET-invA recombinant and expressed. The recombinant InvA protein was purified by metal- chelating affinity chromatography. And the polyclonal anti- InvA fusion protein antibodies were obtained via immunning CD1 mice and the rabbits of New Zealand species with the purified protein. This research will be very helpful for the further study on the characteristics and applifications of the salmonella InvA.Methods:Three parts of research contents were included in this study.1. The construction of Sallmonella invA gene prokaryotic expression recombinants. InvA gene was amplified by PCR from genomic DNA of salmonella and inserted into expression plasmid pET-30c(+) to generate pET-invA. The recombinant plasmid was transformed into the cloning host E.coli DH5a. The positive clones were screened with Kanamycin selection, and the recombinant plasmid was confirmed by double enzyme digestion and sequence analysis.2. Expression purification and confirmation of the recombinant InvA fusion protein. The recombinant plasmid pET-invA was transformed into the expression host E.coli BL21(DE3). The recombinant protein was expressed by the induction of isopropyl-β-D-thiogalactopyranoside (IPTG) and was analyzed by SDS-PAGE. Then the InvA fusion protein was purified by metal-chelating...