Abstract: ObjectiveTo obtain recombinant GlcB protein efficiently expressed in E. coli. MethodsThe M. tuberculosis (M.tb) glcB gene was amplified by PCR and then cloned into plasmid pTA2. After sequencing, the glcB gene was cloned into plasmid pET30a(+), and expressed in E. coli BL21 (DE3). ResultsWhen E. coli BL21 (DE3) containing recombinant plasmid pET30a(+):glcB was induced by 0.4mmol/L of IPTG for 4h, the quantity of recombinant protein expressed in E. coli was maximum. The recombinant protein GlcB with molecular weight 92KD was expressed in incusion bodies of E. coli, and amounted to 50% of total bacterial protein. The purity of the protein purified through the Ni-NTA resin reached 90%. ConclusionsThe prokaryotic expression plasmid pET30a(+):glcB was constructed successfully, and the recombinant protein GlcB was obtained.

Key words: Mycobacterium tuberculosis, Recombinant proteins