Cloning, Expression Of 987P Fimbrial Subunit FasG Gene And Praparation Of Its Monoclonal Antibody

Escherichia coli is one of the most important pathogens in young animals, causing diarrhea,oedema disease or colisepticemia. Infection of enterotoxigenic Escherichia coli (ETEC) is a major cause of diarrhea and death in preweaned pigs.The organism has two types of virulence factors including fimbrial adhensins and enterotoxins. Fimbriae are hair-like proteinaceous appendages, playing an essential role in the pathogenesis of E.coli infection by mediating adhesion to epithelial cells. When the adhesins on the surface of bacteria bind to corresponding acceptor on the surface of host's cells, bacteria are stably adhered to host's cells so that the bacteria can localize in local tissues and produce toxin or destroy tissues, leading to diarrhea.To facilitate investigation into adhesion mechanism of 987P+ETEC, faeG gene of 1119bp was amplified by high fidelity PCR using the DNA template of strain E.coli987.The PCR product was digested by restriction enzymes and then subcloned into the prokaryotic expression vector pET-16b. The (His)10-FasG fusion protein of about 40KDa was expressed in the host cell BL21(DE3) E.coli after by IPTG induction, which was revealed by SDS-PAGE.For acquire the monoclonal antibody against FasG protein, the fusion protein was...