Expression of human sperm protein 17 gene in E. coli

    YANG Aiª²Long, LI Fangª²Qiu, ZHANG Jianª²Qiong, XU Lianª²Hong

    1Genetic Research Center, Medical College of Southeast University, Nanjing 210009, China, 2 PLA Center of Medical Laboratory Sciences, Nanjing General Hospital, Nanjing Military Area Command, Nanjing 210002, China

    ¡¾Abstract¡¿ AIM: To clone human sperm protein 17(hSp17)cDNA and to construct expression vector producing recombinant protein. METHODS: hSp17 cDNA amplified by RTª²PCR from human testis was inserted into plasmid pETª²28a(+) and plasmid pETª²28a(+)/hSp17 transformed E .coli BL21(DE3). Recombinant protein hSp17 was produced under the induction of IPTG, analyzed by SDSª²PAGE, identified by Western blot with specific mAb and purified by Niª²NTA Hisª²Bind resin. RESULTS: The cDNA of hSp17 was cloned and sequenced. Recombinant protein (apparent Mr 27 500) was highly expressed and demonstrated by Western blot. CONCLUSION: hSp17 gene has been successfully cloned and expressed.

    ¡¾Keywords¡¿ human sperm protein 17; cloning, molecular; gene expession; blotting; western ......