Package and identification of replicationª²deficient recombinant adenovirus expression vector of hTERT

    CHEN Ling, XIANG Guoª²Chun, LI Xiangª²Hong, CAI Yongª²Guo, LI Jinª²Jin, FANG Dian Chun, LUO Yuanª²Hui, LI Zhiª²Hong£¬YANG Shiª²Ming

    PLA Gastrointestinal Center, Southwest Hospital, Third Military Medical University, Chongqing 400038, China

    ¡¾Abstract¡¿ AIM: To construct a replicationª²deficient recombinant adenovirus expression vector of hTERT (Adª²hTERT). METHODS: The hTERT gene was cloned at the downstream of CMV promoter of the adenoviral shuttle plasmid pDC315 in sense direction and the resultant recombinant plasmid pDC315ª²hTERT was cotransfected into HEK 293 cells together with plasmid pBHGlox (deltaE1, 3) containing adenoviral genome. The adenovirus expression vector was then obtained and identified by infection test, electron microscopic observation and PCR coª²amplification. RESULTS: After purification and concintration, the titer of Adª²hTERT reached 5¡Á1012 pfu/L. Virus particles could be found in HEK 293 cells under transmission electron microscope. Both adenovirus and hTERT special fragments could be amplified from Adª²hTERT by PCR, whereas hTERT special fragment could not be amplified from the control. CONCLUSION: The replicationª²deficient recombinant adenovirus expression vector of hTERT is successfully constructed. ......