Gene clone and activity assay of apoptin
- VernacularTitle:凋亡素的克隆及诱导HeLa细胞凋亡
- Author:
GuoJing, SUN
;
Xin, TONG
;
ZhiXian, SUN
- Publication Type:Journal Article
- From:Bulletin of The Academy of Military Medical Sciences
2001;25(2):85-88
- CountryChina
- Language:Chinese
-
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.
