Cloning,expression and purification of six Brucella protein

The aim of this study is to construct a prokaryotic expression vector for six brucella proteins,as well as express and purify the recombinant protein.Brucella strain 16M genome sequence was extracted and used as template in polymerase chain reactions to amplify six genes,then the PCR product was cloned into the plasmid pET32a(+) and introduced into the competent cells E.coli BL21(DE3).The protein expression was induced by IPTG and examined by SDS-PAGE and Western blot,then purified by HisTrapTM HP.In this study,six brucella proteins were successfully cloned,the expression condition was optimized,the target proteins were strongly expressed,and highly purified protein were obtained after purified.This study lays a foundation for advance study for immunogenicity and protection of the six brucella proteins.