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 gone was obtained from pGEM3zf-F with Xho Ⅰ and Hind Ⅲ ,cloned into adenovirase shuttle vector pShutth-CMV,and then the resulting pShutth-CMV/F was transformed into E. coli BJS183/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. Conclusion We have obtained a strain of rephcation-defective adenovirus FGAd/F encoding RSV F protein,which can be used further to investigate its protective efficacy against RSV infection in vivo. Key words: Recombinant adenovirus; Respiratory syncytial virus; Fusion glycoprotein