Subcloning and expression of recombinant Echinococcus granulosus antigen B, in Pqe-30 expression Vector

ABSTRACT

Echinococcosis or hydatid disease is a zoonotic infection caused by larval [metacestode] stages of cestodes belonging to the genus Echinococcus, family Taeniidae. We aimed to subclone antigen B gene in pQE-30 plasmid, its expression, and purification. We subcloned HI gene into pQE-30 expression vector. The recombinant vector was transformed into E. coli, M15 and mass cultured. The subcloned gene was expressed by IPTG. Subcloning of gene was confirmed by both PCR and enzyme digestion. Production of recombinant protein was confirmed by SDS-PAGE. Western blot analysis was carried out by both His-Tag monoclonal Ab and human serum to estimate the expressed potein in E. coli cells. Recombinant protein was purified and its specificity was proved by Western blotting. Production of this recombinant protein can increased sensitivity and specificity in serological test [ELISA]