Construction and Identification of adenovirus vector encoding mouse PGC1? gene
- VernacularTitle:编码小鼠PGC1?基因的重组腺病毒载体构建及鉴定
- Author:
Shikun MA
- Publication Type:Journal Article
- Keywords:
PGC1?;
Adenovirus vector;
Homologous recombination
- From:
Journal of Chongqing Medical University
1987;0(01):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To construct an adenovirus vector encoding mouse PGC1? gene for further study on its function. Methods: Recombinant shuttle plasmid pAdTrack-CMV-PGC1? was linearized by PmeⅠ and transformed into Escherichia coli BJ5183 carrying backbone plasmid pAdeasy-1 to obtain adenovirus plasmid through homologous recombination. The recombinant adenovirus plasmid was digested by PacⅠ and transfected into HEK293 cells to form adenvirus particles. The virus titer was determined by TCID50 method. The expression of PGC1? in 293 cells was detected by Western blot. Results: The packaging cell showed typical CPE,and the virus titer was about 3?1010 TCID50/ml after amplification. The expression of PGC1? mediated by the recombinant adenovirus vector was verified by Western blot. Conclusion: The adenovirus vector encoding mouse PGC1? gene was successfully constructed,and the protein was observed in 293 cell.
