Cloning and prokaryotic expression of Mycobacterium tuberculosis Rv2450c gene

    LI Liª²Qing, SU Mingª²Quan, LI Liª²Wen, HAO Xiaoª²Ke

    1Clinical Laboratory, 2PLA Institute of Orthopedics, Xijing Hospital, Fourth Military Medical University, Xi¬ðan 710033, China

    ¡¾Abstract¡¿ AIM: To explore whether the Mycobacterium tuberculosis Rv2450c gene can promote its own resuscitation and growth. METHODS: Mycobacterium tuberculosis was cultured and genome DNA was extracted. Rv2450c gene was obtained by PCR using specific primers, cloned into BamH¢ñ and EcoR¢ñ site of pGEXª²4Tª²2 expression vector and confirmed by sequencing. After the recombinant expression vector being transformed into E.coli BL21(DE3), the recombinant bacteria were induced at 30¡æ for 5 h and the fusion protein GSTª²Rv2450c was analyzed by SDSª²PAGE. RESULTS: DNA sequencing results showed that the Rv2450c gene amplified was exactly consistent with the sequence reported in GenBank and the SDSª²PAGE analysis demonstrated that the Rv2450c was expressed in E.coli BL21(DE3). The protein band amounted to 22.9% of total bacteria protein.CONCLUSION: Mycobacterium tuberculosis Rv2450c gene is successfully cloned and expressed in E.coli BL21(DE3). ......