Construction and identification of replication deficient recombinant adenovirus encoding F gene of subgroup A human respiratory syncytial virus.
- Author:
Yuan YUAN
1
;
Jin-sheng HE
;
Yuan-hui FU
;
Mei ZHANG
;
Qian TANG
;
Dong-liang LI
;
Wei WEI
;
Jian-guo QU
;
Tao HONG
Author Information
- Publication Type:Journal Article
- MeSH: Adenoviridae; genetics; Adenoviridae Infections; genetics; Cloning, Molecular; Gene Expression; Genetic Vectors; genetics; metabolism; Humans; Recombinant Fusion Proteins; genetics; metabolism; Respiratory Syncytial Virus, Human; genetics; Respiratory Syncytial Viruses; genetics; Viral Fusion Proteins; genetics; metabolism; Virus Replication
- From: Chinese Journal of Experimental and Clinical Virology 2008;22(6):428-430
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVEA strain of replication deficient recombinant adenvirus encoding fusion glycoprotein (F) of subgroup A human respiratory syncytial virus (RSV) was constructed and the expression of F was identified.
METHODSThe F gene was obtained from pGEM3zf-F with Xho I and Hind III, cloned into adenoviruse shuttle vector pShuttle-CMV,and then the resulting pShuttle-CMV/F was transformed into E. coli BJ5183/p with pAdeasy-1 to produce pre-adenoviral plasmid encoding F by homologous recombination. This resultant plasmid was linearized by digestion with Pac I and transfected into 293 packaging cells to generate FGAd-F. Finally, the expression of F protein was identified by Western Blot analysis.
RESULTSFGAd/F was successfully constructed, and the expression of RSV F protein was identified by Western Blot.
CONCLUSIONWe have obtained a strain of replication-defective adenovirus FGAd/F encoding RSV F protein, which can be used further to investigate its protective efficacy against RSV infection in vivo.
