[Construction and identification of replication deficient recombinant adenovirus encoding F gene of subgroup A human respiratory syncytial virus].
OBJECTIVE
A 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.


METHODS
The 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.


RESULTS
FGAd/F was successfully constructed, and the expression of RSV F protein was identified by Western Blot.


CONCLUSIONS
We 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.
Yuan
Yuan
Y
Department of Immunology, Anhui Medical University, Hefei 230032, China.
He
Jin-sheng
JS
Fu
Yuan-hui
YH
Zhang
Mei
M
Tang
Qian
Q
Li
Dong-liang
DL
Wei
Wei
W
Qu
Jian-guo
JG
Hong
Tao
T
D000256:Adenoviridae / Q000235:genetics*; D000257:Adenoviridae Infections / Q000235:genetics; D003001:Cloning, Molecular; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D006801:Humans; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D018113:Respiratory Syncytial Virus, Human / Q000235:genetics; D012136:Respiratory Syncytial Viruses / Q000235:genetics; D014760:Viral Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D014779:Virus Replication
D011993:Recombinant Fusion Proteins; D014760:Viral Fusion Proteins