Construction Of Recombinant Plasmid Of Hantaan Virus Truncated S Genome Segment And Its Expression In C.perfringens

Objective To construct a recombinant plasmid containing truncated region of S genome segment of Hantaan virus, and made it express in C. perfringens which is a vector of oral vaccine.Methods Target gene-SA was obtained by PCR from pGEX-4T-2/SA , and cloned into shuttle vector pJRC200. The recombinant plasmid pJRC200-SA was identified by restriction enzyme analysis, PCR, DNA sequencing, and transformed into C. perfringens by electroporation. The C. perfringens was cultured in fluid thioglycolate broth in an anaerobic condition, and sporulation broth was used to obtain sporulating cultures of C. perfringens. The C. perfringens spore was handled with ultrasonic crasher , and the expression of protein was detected by SDS-PAGE and western blotting .Result Restriction enzyme analysis, PCR, and DNA sequencing demonstrated that the truncated region of S genome segment of hantaan virus was successfully inserted into pJRC200. Western blotting analyses demonstrated that the molecular weights of SA was 27000 Da, and presented specific antigenicity .Conclusion The plasmid pJRC200-SA was successfully constructed, and expressed in C. perfringens , which lay a foundation for the research of oral vaccine.