Establishment of Murine Tumor Cell Line Expressing HPV types 6b and 11 E6/E7 Genes
- VernacularTitle:表达人乳头瘤病毒6b和11型E6/E7基因的小鼠肿瘤细胞株的建立
- Author:
Kejia ZHAO
;
Hao CHENG
;
Minli CHEN
;
Zhishan DING
;
Liyi GENG
;
Yongming FANG
- Publication Type:Journal Article
- Keywords:
Papillomavirus, human;
Gene expression;
Plasmids;
Transfection
- From:
Chinese Journal of Dermatology
1994;0(02):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct four expression plasmids, pcDNA3.1-GFP/HPV6bE6, pcDNA3.1-GFP/HPV6bE7, pcDNA3.1-GFP/HPV11E6, pcDNA3.1-GFP/HPV11E7 and their transfected murine cell lines. Methods The Four recombinant expression plasmids comprising HPV6bE6,HPV6bE7,HPV11E6 and HPV11E7 linked with GFP, respectively, were constructed and transfected to B16 cells by lipofectamine kit. Positive clones were selected by G418 and observed by fluorescent microscopy and identified by RT-PCR. Results The four constructed recombinant plasmids were authenticated by restriction enzyme digestion and DNA sequencing. Under the fluorescent microscope, the green fluorescence could be observed in cytoplasm and nucleus of four transfected B16 cell lines. The RNA extracted from positively transfected clones resistant to G418 were analyzed by RT-PCR, which demonstrated the presence of four expected fragments. Conclusions The transfected murine cell lines B16 can express HPV6bE6,HPV6bE7,HPV11E6 and HPV11E7 gene. These transfected cell lines can be further transplanted to mice in order to investigate the biological properties and immunological mechanisms of these genes in vivo.
