我国部分地区鸡新城疫病毒流行株遗传变异分析  

Genetic Variation Analysis of Chicken-origined NDV Strains Recently Isolated From Some Areas in China

作　　者：陆广富[1,2] 吴卫东[2] 张建军[2] 葛兆宏[1] 王建业[2] 王永坤[2] 朱国强[2] 

机构地区：[1]江苏畜牧兽医职业技术学院,江苏泰州225300 [2]扬州大学兽医学院,江苏扬州225009

出　　处：《中国家禽》2010年第13期23-26,共4页China Poultry

基　　金：国家自然科学基金(30571374;30771603);江苏省属高校自然科学重大基础研究项目(08KJA230002)

摘　　要：将近年来从不同地区临床病例中分离鉴定的10株鸡新城疫病毒(NDV)流行株,用RT-PCR方法分别扩增出大小为535 bp的F基因重要功能区,并克隆到pGEM-T载体上,经酶切分析结合核苷酸序列测定而确诊。10株分离株核苷酸序列同源性达到81.9%～99.4%,其中8个分离株F蛋白裂解位点的氨基酸顺序为:112R-R-Q/R-K/R/R/F117,为NDV强毒株,且具有基因Ⅶ型的典型特征,即在101位和121位氨基酸残基分别为K(赖氨酸)和V(缬氨酸);其余2株病毒F蛋白裂解位点的氨基酸顺序为:112G-R-Q-G-R-L117,表明它们为NDV弱毒株,在13位和17位氨基酸分别为M(蛋氨酸)和I(异亮氨酸),属于基因Ⅱ型。根据上述测序结果并参照已发表NDV F基因序列绘制出遗传进化树,遗传进化树显示,上述8个强毒株和台湾95年分离株同源性较高,而2个弱毒株与La Sota株同源性较高。Ten strains of Newcastle disease virus （NDV）were recently isolated and identified from the sick chickens infected with NDV. Main function segment of fusion （F） gene with the length of 535 bp for the above 10 isolates were amplified by RT-PCR method,then cloned into pGEM-T vector. The positive recombinants were confirmed by the combination methods of both restriction enzyme digestion and sequencing, respectively. Based on the sequence data for 10 strains,they shared homologue between 81.9% to 99.4% at the nucleic acid level. Eight of all isolates,the amino acids pattern in the F cleavage sites were 112R-R-Q-K-R-F117 with the same of NDV velogenic strain,they also had the characteristic of genotype Ⅷ with amino acids residue pattern of Lysine and Valine at the position 101 and 121,respectively. The other two strains had the same pattern of F cleavage sites as NDV lentogenic strain with amino acids residue pattern of U2G-R-Q-G-R-LU7,and had the characteristic of genotype I1 with amino acids residue pattern of Methionine and Isoleucine at the position 13 and 17,respectively. A phylogenetic tree was drew according to the sequence information among the isolates and the known reference NDV sequences from GenBank. The phylogenetic analysis showed that the eight velogenic strains had a high homologue with Taiwan-origined strain 1995,the other two lentogenic strains had a high homologue with La Sota strain.

关 键 词：鸡 新城疫 腿因 基因Ⅶ型 

分 类 号：S968.313[农业科学—水产养殖]