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

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 cis-direction or trans-direction 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(pDC315-shTERT),while three fragments which lengthened 1.0,2.5 and 3.9 kb were formed in antisense vector(pDC315-ahTERT).Electrophoresis results were completely consistent with theoretical calculation.The DNA sequence measurement confirmed the linking direction of sense and antisense pDC315-hTERT.CONCLUSION: The sense and antisense hTERT adenovirus expression vectors are successfully constructed.