鹅源新城疫病毒F和HN基因的表达及功能鉴定

Expression and functional identification of F and HN gene from Newcastle disease virus of goose origin

鹅源新城疫病毒的F和HN基因经RT-PCR方法从尿囊液中扩增后分别克隆进pCR2．1载体,再分别亚克隆到真核表达载体pCI-neo上,构建了F基因的表达载体(pCI-F)和HN基因的表达载体(pCI-HN)。间接免疫荧光(IFA)鉴定结果表明:F和HN基因均能在细胞中得到较好的表达;将pCI-F和pCI-HN共转染CEF和COS细胞,转染的细胞中能形成明显的合胞体。而用pCI-F单独进行转染时,不能产生合胞体,说明F蛋白要发挥其融合作用必须要有HN蛋白的参与,同时也进一步证明F和HN蛋白与病毒的融合特性密切相关。

F and HN gene of Newcastle disease virus of goose origin were amplified and cloned into pCR2.1 vector. F and HN gene from pCR2.1 vector were subcloned into eukaryotic expression vector, pCI-neo, to construct pCI-F and pCIHN respectively. The positive clones were identified by enzyme digestion, PCR and sequencing. The results of IFA showed that F and HN gene expressed well in COS-1 cell. The experiment of cotransfection with pCI-F and pCI-HN revealed that cO-expression of pCI-F and pCI-HN could induce syncytia formation on transfected cells, but the cells transfected with pCI- F alone couldn＇t form syncytia. The results mentioned above confirmed that F and HN protein were associated with the fu- sion function of NDV.