Construction and identification of mouse Osf2/Cbfal gene recombinant adenovirus
- VernacularTitle:小鼠Osf2/Cbfa1基因的重组腺病毒构建及鉴定
- Author:
Shiwu DONG
;
Dajun YING
;
Guangjiu LIU
;
Chuhong ZHU
;
Wei ZHANG
- Publication Type:Journal Article
- Keywords:
adenovirus;
Osf2/Cbfal gene;
homogenous recombination
- From:Journal of Third Military Medical University
2003;0(21):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct the recombinant adenovirus of mouse Osf2/Cbfal gene and to observe its ability to infect NIH3T3 fibroblasts. Methods The Osf2/Cbfal gene fragment was cloned into the shuttle plasmid pAdTrack-CMV to form the transfer vector by the method of homogenous recombination in bacteria. Then the recombinant adenovirus was transfected into NIH3T3 cells using Lipofectine DOTAP, The target gene was detected by poly-merase chain reaction (PCR). The titer and its infection rate were determined using the green fluorescent protein (GFP) expression in the shuttle plasmid. Results Restriction endonuclease and PCR analyses confirmed that the Osf2/Cbfal gene was successfully inserted into the adenovirus vector. The titer of the recombinant adenovirus was 1.6?1012 pfu/ml. The adenovirus had a strong effect on NIH3T3 cells. Conclusion The recombinant adenovirus containing Osf2/Cbfal gene was successfully constructed by the method of homogenous recombination in bacteria.
