羊痘病毒江苏株P32基因的克隆与序列分析  

Cloning and Sequence Analysis of P32 Gene of Capripox Virus

作　　者：张志成[1] 陈琼 庞银芳 陆静静[1] 戴薇[1] 张大淦[1] 陈钟鸣[1] 

机构地区：[1]金陵科技学院动物科学与技术学院,江苏南京210038 [2]江苏省丹阳市吕城兽医站,江苏丹阳212351

出　　处：《金陵科技学院学报》2010年第4期79-83,共5页Journal of Jinling Institute of Technology

基　　金：金陵科技学院科研基金(Jit-n-2009-011);南京市教育科学研究"十一五"规划课题(L09/002)

摘　　要：根据GenBank中羊痘病毒基因（CPV）的核苷酸序列,设计了1对特异性检测引物。采用此引物检测来自于江苏丹阳的24份发病羊的皮肤痂皮。参考已发表的P32全基因的特异性引物,随机选择4份阳性DNA进行P32片段的扩增。将扩增产物纯化后连接pMD18-T载体,转化DH5α大肠埃希氏菌,选择阳性克隆送基因公司测序。应用DNAStar等分子生物学软件,对所测P32序列与GenBank中的国内外CPV毒株进行了同源性比较和遗传进化树分析。结果发现,24份疑似病料检测全是阳性;4个P32基因扩增产物均为1 023 bp,且样品间P32基因的核苷酸同源性为100%,与GenBank上已发表的的P32基因序列的相似性为97.7%~100%,且中国分离株在进化上属于同一分支。According to the nucleotide sequences of Capripoxvirus（CPV） in GenBank,a pair of specific primers is designed and detected from 24 crust skins of pathogenetic sheep samples isolated from Danyang,Jiangsu Province by PCR.Referring to the published P32 entire gene-specific primers,4 positive DNA samples are randomly selected to be amplified.Amplified products are purified and cloned into the pMD-18 T vectors,then transformed into Escherichia coli DH5α and the positive recombinant plasmids are sequenced.Based on homology comparison and evolutionary tree analysis of P32 genome sequences with CPV strains isolated from home and abroad in GenBank by DNAStar software,the results demonstrate that 24 suspected samples are all positive;the size of 4 amplification products is 1023bp,the P32 gene nucleotide homology among samples is 100%,which has the similarity of 97.7% to 100% with the P32 gene sequence published in GenBank,and the Chinese isolated strains belong to the same branch in evolution.

关 键 词：羊痘病毒 P32基因 克隆 序列分析 

分 类 号：S852.654[农业科学—基础兽医学]