猪瘟病毒石门株特异cDNA片段的扩增与序列分析  

SPECIFIC SEQUENCE AMPLIFICATION OF CHINESEHOG CHOLERA VIRUS STRAIN SHIMEN

作　　者：涂长春[1] 李红卫[1] 金扩世[1] 章金钢[1] 白安斌[1] 刘士英[1] 扈荣良[1] 殷震[1] 

机构地区：[1]长春农牧大学兽医研究所,广西兽医研究所

出　　处：《病毒学报》1994年第1期33-38,共6页Chinese Journal of Virology

基　　金：国家"863"生物技术领域资助课题。

摘　　要：以猪瘟病毒感染细胞中提取的细胞总ＲＮＡ为模板，应用反转录─聚合酶链反应，在化学合成的两对特异引物引导下，成功地扩增出了两个石门株的ｃＤＮＡ片段。电泳证明它们的大小与预计的３４６ｂｐ和１２０ｂｐ完全一致。酶切分析证实了它们应有的酶切位点。随后对这两个片段进行了克隆和序列测定，并与国外株的序列进行了比较。结果证明本实验扩增的两个ｃＤＮＡ序列与Ａｌｆｏｒｔ株和Ｂｒｅｓｉａ株的同源性分别是９５．２％和９８．５％。Based on the cDNA sequence of hog cholera virus (HCV)strain Alfort ,2 pairs of prim-ers were chemically synthesized and used to amplify 2 specific sequences of Chinese HCV strainShimen by reverse transcription-polymerase chain reaction(RT-PCR)from total intact intact RNAextracted from infected cell culture.Expected Size of 346 and 120 base pairs of 2 fragments and their specificity were confirmed by restriction endonuclease digestion,and they were cloned and sequenced.The sequencing data were compared with those previously published and used to deduce genetic relationship between the HCV strains .The result showed that the sequenced region of Shimen strain had high degree of homology with Brescia (98.5%)and Alfort (95.2%),and that strain Brescia lacks cytosine base at 252 position on 5’non-coding region as compared with that of Alfort and Shimen.

关 键 词：猪瘟病毒 CDNA 扩增 序列分析 

分 类 号：S852.651[农业科学—基础兽医学]