Construction of Baculovirus Expression Vector of Human Insulin Gene and Its Expression in Sf_9 Cells

The transpositional recombinant vector was constructed firstly. Then it was transformed in E. coli DH10Bac and cultured in LB plate which contained Gm, Kan, X gal and IPTG. The white bacterial colonies were positive recombinant Bacmid named as Bacmid/mINS.The recombinant baculoviruses were obtained after the transfection of the recombinant vector into Sf 9 lines. The techniques of Tricine SDS PAGE and Western blotting were used to detect and identify the products expressed in Sf 9 lines by recombinant baculovirus. The results indicated that the products expressed in Sf 9 lines by baculovirus vector system were most likely proinsulin.None of the proinsulin and insulin were detected in the supernatant of Sf 9 cell culture.