Prokaryotic expression of human UROC28 gene and preparation of its polyclonal antibody

    WANG Chunª²Yang, YANG Liª²Jun, WU Yuanª²Ming, SHAO Guoª²Xin, WANG He, CHEN Nanª²Chun, CHEN Suª²Min

    1Department of Urology, Xijing Hospital, 2Department of Biochemistry and Molecular Biology, School of Basic Medicine, Fourth Military Medical University, Xi¡¯an 710033, China

    ¡¾Abstract¡¿ AIM: To express human UROC28 gene and to prepare its specific polyclonal antibodies for the diagnosis and treatment of prostate cancer. METHODS: UROC28 gene was cloned into the expression vector pRSETª²c and Hisª²UROC28 fusion protein was expressed by IPTG induction and purified with SDSª²PAGE from the total proteins of E.coli BLª²21 (DE3). New Zealand rabbits were immunized with this protein to prepare polyclonal antiserum and the specificity of the polyclonal antiª²UROC28 antibody was determined by Western blot. RESULTS: Hisª²UROC28 fusion protein (Mr 22 000) was successfully expressed. Western blot showed that the polyclonal antiª²UROC28 antibody recognize native UROC28 and was of high specificity against UROC28 without crossª²reaction. The expression of UROC28 was also successfully detected in prostate cancer tissues. CONCLUSION: UROC28 has been successfully expressed in BLª²21. Polyclonal antiª²UROC28 antibody with high specificity has been prepared and applied successfully as a tumor marker. ......