牦牛FMDV持续感染分离株非结构蛋白P2的克隆及基因特征分析.pdfVIP

甘肃事业大学学掘 蛐 42 嚣 2007 年 10 月 JOURNAL OF GANSU AGR1CULTURAL UNIVERSITY ，jI(月刊 第 5 期 16-19 把牛 FMDV 持续感染分离株非结构蛋白白的 克隆及基因特征分析 魏小娟1 ， 2 ，常惠芸2 ，张继瑜1 ，丛国政2 ，唐江山2 【1.中国现业科学院兰州畜牧与兽药研究所，甘肃兰州 730050 ,2 中国最业 科学院兰拥兽医研究所，甘肃兰删 730046) 摘要z 以 FMDV 持续感阜模型葫物乾牛的食遭/咽部分离物(O/P 液)为反转量模植，用 1 对特异性引物扩增 目的 cDNA.然后与 pMD18一T 载体连接井转化JM109 菌棒，再经重组质植电睡、PCR 和 ECQRI 酶切鉴定，序列 测量和分析结果量明，分离峙与 Cbinal阳的同摞性最高达 93.4 %.而与 Akesu/58 序列的同源性为 90.3 坷 s 推导 的l!;基酸序列同IJ.性分别为 97.8% 、 96.7 %. 9是键胃: FMDV，持续感阜，P2 非结构蛋白s 克隆a 序列分析 中图骨费号: 5 852.659.6 ,Q 786 立献标识码:A 宜章躏号: 1003-4315(2007) 05-0016-04 岛10lecular characterization of non-structural protein P2 of FMDV in yak persistent infection strain WEI Xiao-juan .CHANG Hui-yun ,ZHANG Ji-yu •CONG Guo-zheng •TANG Jiang-shan (1. Lanthou lnstltute of Anìmal and VeterÌnary Pharmaceutìcs Sciences ,Chìnese Acaðemy 01 Agriculturøl 坠ien四.. Lan曲OU 730050.Chir施I 2. Lanzhou Veterinary Institu饵.Chínese A国demy of Agricultural Sciences ,Lanzhou 730046 ,China) Abstract , Using oesophagea!-pharyngea!O/Pl f!uid that iso!ated from FMDV A祉esu/58 persistent in- fection model yak as templet to reverse transcript ,amplify its P2 tract with a paÌr of primers. The amplified cDNA products were cloned into pMD18一T vectors and transformed into JMI09 ,the recombinant plas- mids were identified by electrophoresis ,EcoR 1 cleavage and PCR. The nucleotide sequence of the expected P2 tract was determined by SangerS DNA sequence method ,and amino acid sequence was compared with the P2 tracts of the other seven FMDV strains. It was shown that the homology of P2 tract of isolation strain with China/9