Construction of the prokaryotic expression vector with Rv0901 of Mycobacterium tuberculosis and its expression in E.coli

Objective To construct prokaryotic expression vector as the basis for exploring the function of a unknown gene Rv0901 of Mycobacterium tuberculosis.Methods The gene encoding Rv0901 protein of Mycobacterium tuberculosis H37Rv strain was amplified by PCR and cloned into prokaryotic expression vector pGEX-1λT.The recombinant plasmid was transformed into E.coli JM109 to express the fusion protein GST-Rv0901 induced by IPTG.The expression product was analyzed by SDS-PAGE and purified by the Bulk GST Purification Module.Results The gene Rv0901 was amplified accurately from the genome DNA of H37Rv.A recombinant fused expression vector pGEX-Rv0901 was constructed.The recombinant plasmid can express the fusion protein GST-Rv0901 stably.Conclusions The 45 kDa fusion protein GST-Rv0901 was successfully expressed and purified.It provided the basis for the further study of the gene Rv0901.