鸡新城疫病毒ND-xx08株F基因的克隆及分析  

Cloning and Analysis of F Gene from Newcastle Disease Virus ND-xx08 Isolate

作　　者：王选年[1,2] 贾文科[3] 银梅[1] 岳峰[1] 田献礼[1] 

机构地区：[1]新乡学院生命科学与技术系,河南新乡453003 [2]河南科技学院动物科学学院,河南新乡453003 [3]河南农业大学牧医工程学院,郑州450002

出　　处：《新乡学院学报》2010年第1期43-46,共4页Journal of Xinxiang University

基　　金：河南省高校科技创新团队支持计划(2008IRTSTHN001)

摘　　要：新城疫病毒（NDV）ND-xx08毒株经10 d龄SPF鸡胚增殖后,提取其基因组RNA并反转录成cDNA,用NDV F基因特异性引物,经PCR扩增后获得与F基因预期大小一致的DNA片段。将NDV F基因片段克隆到pMD18-T载体上,并进行EcoR I和Hind III双酶切鉴定和测序鉴定。结果显示,ND-xx08毒株F基因片段的长度为1 662 bp,共编码554个氨基酸,F蛋白的裂解位点为112R-R-Q-K-R-F117,是典型强毒株氨基酸序列结构。将NDV ND-xx08株F基因的47 bp到420 bp序列与新城疫病毒基因型I至基因型Ⅸ毒株的相同序列绘制病毒基因进化树,显示ND-xx08分离株属于基因Ⅶe型。将NDV ND-xx08株F全基因与国内外发表的23株NDV F基因核苷酸序列和氨基酸序列的同源性比较分析,结果表明,其核苷酸序列的同源性在82.7%~97.8%之间,氨基酸同源性在87.5%~97.7%之间。Newcastle disease virus（NDV） RNA was isolated directly from ND-xx08 isolate,which propagated by inoculating in SPF embryonated eggs.The viral RNA was reverse transcribed into cDNA and then NDV F gene DNA fragment was cloned into pMD-18 vector by F gene spicific primer pair amplification for sequence and double enzyme digested by EcoR I and Hind III identification.The full length F gene comprised 1662 bp which encodes 553 amino acids.The deduced amino acid sequence of the ND-xx08 isolate F protein at the cleavage site was 112RRQKRF117,which is typically found in virulent NDV isolates.Phylogenetic analysis revealed that the isolate belonged to genotype VIIe by compared the ND-xx08 isolate with other NDV isolates representative of genetic groups I to IX based on the F gene nucleotide sequence at position 47 to 420（374 bps）.Results showed that the F gene nucleotide sequence homology was 82.7% to 97.8% and amino acid homology was 87.5% to 97.7% compared with the 23 published NDV sequences.

关 键 词：鸡 新城疫病毒 ND-xx08毒株 F基因 

分 类 号：S855.3[农业科学—临床兽医学]