Cloning and Fusion Expression of esat6-cfp10 of Mycobacterlum Tuberculosis and Its Purification

To construct a recombinant plasmid carrying esat6 and cfp10 of mycobacterium tuberculosis and express this fusion protein, cfp10 gene was amplified by PCR with specific primer from genome of MTB H37Rv. cfp10 was inserted to the cloning vector pGEM-7ZF( +) with esat6 gene. Then esat6-cfp10 digested by restriction enzyme, it was cloned to expressing vector pPROEX?HTb and expressed in E. coli DH5α. SDS-PAGE showed that a 28 kD fusion protein was expressed. Purified proteins was obtained via Ni-NTA purification system. Western blot suggested that this fusion protein has a good immu-nogenicity.