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

VernacularTitle:人呼吸道合胞病毒融合糖蛋白非复制型重组腺病毒的构建和表达
Author: Yuan YUAN ¹ ; Jin-Sheng HE ; Yuan-Hui FU ; Mei ZHANG ; Qian TANG ; Dong-Liang LI ; Wei WE ; Jian-Guo QU ; Tao HONG
Author Information

1. 安徽省疾病预防控制中心
Keywords: Recombinant adenovirus; Respiratory syncytial virus; Fusion glycoprotein
From: Chinese Journal of Experimental and Clinical Virology 2008;22(6):428-430
CountryChina
Language:Chinese
Abstract: 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.