Gene clone and activity assay of apoptin

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. [