Cloning and expression of human survivin gene

    GUO Wanª²Hong, CHENG Shiª²Yin, ZHANG Huiª²Zhong, LIN Fang

    1Department of Otolaryngology, 2Department of Clinical Central Laboratory, Tangdu Hospital, Fourth Military Medical University, Xi¡¯an 710038, China

    ¡¾Abstract¡¿ AIM: To clone, express and identify human survivin gene. METHODS: Total RNA was extracted from human Hepª²2 laryngocarcinoma cells and the whole length gene of survivin was obtained by RTª²PCR. The survivin gene was cloned into pGEMª²Teasy vector and sequenced. Then the gene was inserted to XhoI and NdeI site of pRSETª²c expression vector to construct the expression vector, which was transformed into E.coli BL21. After the transformed bacteria were induced at IPTG for 4-5 h, the expressed protein was analyzed by SDSª²PAGE. RESULTS: DNA sequencing results showed that survivin gene was exactly consistent with the sequence reported in GeneBank. SDSª²PAGE analysis demonstrated that survivin protein was expressed in E.coli and the relative molecular mass of it was 16.5¡Á103. The protein band amounted to 30% of total bacteria protein. CONCLUSION: The results show that survivin gene has been successfully cloned and expressed. It lays a foundation for further studies of the applications of survivin in the tumor diagnosis and therapy. ......