Construction and identification of recombinant rBCG-Rv2029c vaccine of Mycobacterium tuberculosis / 中国人兽共患病学报

ABSTRACT

We constructed a recombinant vaccine of Mycobacterium tuberculosis rBCG-Rv2029c,and then identified it.Rv2029c antigen encoding gene was amplified by PCR.The enzyme digestion products were ligated into rpMV261-2029c recombinant plasmid,after double digestion of Rv2029c and pmv261 vector,and then we introduced the plasmid into BCG to construct rBCG-Rv2029c recombinant vaccine by electroporation method.Finally,we analyzed the expression of the recombinant protein by SDS-PAGE and Western blotting.A total of 1 020 bp Rv2029c gene successfully amplified by PCR was inserted into the plasmid pmv261,then the fusion gene was successfully transduced into BCG.After identified by double enzyme digestion,confirmed by gene alignment and by thermally induced with Western blotting,the recombinant protein had a free primary.The recombinant live vaccine of M.tuberculosis rBCG-Rv2029c is successfully constructed,which lay a foundation for the study of the immune mechanism of recombinant vaccine.