Construction and identification of sense and antisense human telomerase catalytic subunit adenovirus expression vector

    CHEN Ling, YANG Shiª²Ming, CAI Yongª²Guo, FANG Dianª²Chun, LUO Yuanª²Hui

    Department of Gastroenterology, Southwest Hospital, Third Military Medical University, Chongqing 400038, China

    ¡¾Abstract¡¿ AIM: To construct an adenovirus expression vector of sense and antisense human telomerase reverse transcriptase (hTERT) gene. METHODS: The hTERT cDNA fragment contained in the pGRN145 vector was cloned into the adenovirus expression vector pDC315 in cisª²direction or transª²direction using DNA recombinant technology. The recombinant vectors were identified by digestion of BamH I and the sense and antisense recombinant vector was further identified by DNA sequencing. RESULTS: After digested by BamH I, two fragments which lengthened 1.0 and 6.4 kb were formed in sense recombinant vector (pDC315ª²shTERT), while three fragments which lengthened 1.0, 2.5 and 3.9 kb were formed in antisense vector (pDC315ª²ahTERT). Electrophoresis results were completely consistent with theoretical calculation. The DNA sequence measurement confirmed the linking direction of sense and antisense pDC315ª²hTERT. CONCLUSION: The sense and antisense hTERT adenovirus expression vectors are successfully constructed. ......