辽宁地区高致病性蓝耳病病毒N基因的克隆及序列分析  

Clonning and Sequential Analysis of N Gene of Liaoning Highly Pathogenic PRRSV Separated Strain

作　　者：刘金玲[1] 魏澍[2] 支海兵 

机构地区：[1]沈阳农业大学畜牧兽医学院,沈阳110866 [2]辽宁省动物疫病预防控制中心,沈阳110164 [3]国兽医药品监察所,北京100081

出　　处：《沈阳农业大学学报》2010年第4期481-483,共3页Journal of Shenyang Agricultural University

基　　金：国家科技攻关项目(413010209);沈阳农业大学校青年基金项目(200705)

摘　　要：从辽宁的本溪、沈阳、抚顺、鞍山等地采集疑似高致病性蓝耳病病料,将采集的肺和淋巴结分别研磨,离心并制备成病毒悬液,用试剂提取RNA,将提取出的RNA反转录成cDNA,然后用特异性引物对目的片段进行克隆扩增,最后将所得目的片段测序并与国内其他分离株进行序列分析。结果获得480bp特异性高致病性蓝耳病病毒N基因片段,与国内其他地区高致病性蓝耳病病毒分离株的N基因相比较,核苷酸序列同源性在91.1%以上,氨基酸序列同源性在98.6%以上,表明高致病性蓝耳病病毒辽宁地区分离株与国内其它分离株亲源关系较近,属于同一分支。The suspect highly porcine reproductive and respiratory syndrome virus sickness materials were collected from Benxi,Shenyang,Fushun,Anshan,etc.in Liaoning.The lung and lymph node were ground separately and centrifugalized for preparing the virus suspension.RNA was extracted with the reagent of reversesly transcripted to cDNA.The target fragment was cloned cond amplified with the specificity primer.The obtained target fragment was conducted for the sequential analysis and compared with other domestic separate strains.As a result we obtained 480bp specificity N gene fragment of highly porcine reproductive and respiratory syndrome virus.Compared with the other domestic highly porcine reproductive and respiratory syndrome virus isolation of N gene,the nucleotide sequence homology was as high as 91.1%,amino acid homology of N protein was above 98.6 %,indicating that there is no significant difference between the viral N gene of Liaoning separated strains and the other domestic strains.therefore the belong to the same cluster.

关 键 词：猪高致病性蓝耳病毒 N基因 克隆 序列分析 

分 类 号：S858.28[农业科学—临床兽医学]