Expression of THANK gene in SMMC-7721 cell line mediated by THANK adenovirus
- VernacularTitle:腺病毒介导的THANK基因在SMMC-7721细胞中的表达
- Author:
Dong WU
;
Feng SHEN
;
Mengchao WU
- Publication Type:Journal Article
- Keywords:
THANK;
gene expression;
liver neoplasms;
adenovirus
- From:
Academic Journal of Second Military Medical University
1985;0(06):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To construct a THANK adenovirus vector and observe expression of THANK in SMMC-7721 cell line after THANK adenovirus infection. Methods: Human THANK cDNA was cloned into shuttle vector pAdTrack-CMV, and the resultant plasmid was linearized and subsequently cotransformed into E.coli BJ5183 with an adenoviral backbone plasmid pAdEasy-1.Recombinants were then selected and the linearized recombinant plasmid was transfected into 293 cell lines. Finally, THANK recombinant adenovirus production was observed by fluorescent microscope and confirmed by PCR and Western blot analysis. Further THANK adenovirus were used to infect SMMC-7721 cells with different MOI. The expression of THANK in SMMC-7721 cells was observed by fluorescent microscope and by PCR and Western blot analysis. Results: A human THANK recombinant adenovirus vector was constructed and infected SMMC-7721 cell line with high efficiency. The expression of THANK in SMMC-7721 cells was over 35 ng/ml 5 d after transfection. Conclusion: Adenovirus can induce stable expression of THANK in SMMC-7721 cells.
