Gene clone and activity assay of apoptin / 军事医学科学院院刊

ABSTRACT

Objective:To obtain apoptin gene and to induce tumor cell (HeLa) apoptosis. Methods:Apoptin gene was amplified by PCR from CAV TK5803 genomic DNA,and was then cloned into pCDNA3.1/His/Topo vector. After confirming by DNA sequencing, apoptin gene was subcloned into pCIneo vector.Recombinant plasmid pCDNA3.1/His/Topo-apoptin and pCIneo-apoptin were transformed into HeLa cells by Lipofectamine reagent. Three days after transfection,the HeLa cell was stained by Honchest 33258 and apoptosis was analysed by fluorescence microscope. Results and Conclusions: The full-length apoptin gene was cloned by PCR and inserted into pCDNA3.1/His/Topo vector. Sequence analysis demonstrated that it was same as published apoptin gene sequence. Three days after transfection, HeLa cell turned into apoptosis.