Construction of prokaryotic recombinant plasmid encoding Mtb8.4 gene of M.tuberculosis and identification

To construct prokaryotic expression recombinant plasmid encoding Mtb8.4 gene of M.tuberculosis. The gene encoding protein Mtb8.4 was amplified by PCR technique. Clone the gene into the downstream of the T7 promoter pET-His, then transformed into E.coli TOP10F′. Positive clone was screened using double digestion and polymerase chain reaction, and farther identified by nucleotide sequence. Mtb8.4 gene was successfully inserted into the prokaryotic plasmid pET-His. The reading frame and gene sequence of recombinant plasmid were correct by sequencing reaction. [Conclusions] Recombinant plasmid pET-His-Mtb8.4 is successfully constructed, It will provide further study for the expression Mtb8.4 protein and its immunological effect, and a potential candidate for inclusion in a subunit vaccine against tuberculosis(TB).