猪瘟病毒石门株与兔化弱毒株gp55基因的克隆与序列分析  

CLONING AND SEQUENCE ANALYSIS OF E1 GENE OF TWO CHINESE HOG CHOLERA VIRUS STRAINS

作　　者：李红卫[1] 涂长春[1] 金扩世 王新平 殷震[1] 

机构地区：[1]中国人民解放军农牧大学军事兽医研究所

出　　处：《病毒学报》1998年第3期257-261,共5页Chinese Journal of Virology

基　　金：国家自然科学基金

摘　　要：猪瘟病毒石门系强毒株及兔化弱毒疫苗株（ＨＣＬＶ株）是我国的标准毒株，囊膜糖蛋白ｇｐ５５（又称Ｅ１）是该病毒最重要的保护性抗原。本实验采用反转录ＰＣＲ扩增了这两个毒株的ｇｐ５５基因。序列分析结果表明：１．石门株和兔化弱毒株糖蛋白Ｅ１的氨基酸序列含有１５个半胱氨酸残基（Ｃｙｓ），其数量及位置与国外４株猪瘟病毒（Ａｌｆｏｒｔ、Ｂｒｅｓｃｉａ、ＡＬＤ和ＧＰＥ－）完全一样。Ｅ１中Ｃｙｓ的数量及位置的保守性说明，该病毒这一保护性抗原在各毒株之间具有相同的骨架结构。２．Ｅ１糖蛋白Ｎ端的信号肽序列及Ｃ端的跨膜区序列是比较保守的，然而位于Ｎ端的抗原区序列是非保守的，其中氨基酸变异较大的区域大约在７０２～７４２氨基酸残基之间。３．所测得的ＨＣＬＶ株Ｅ１的氨基酸序列，与国外测得的ＨＣＬＶ株Ｅ１的氨基酸序列有明显差异，这一差异均一地分布于整个Ｅ１序列。因此，推测国内外同出一宗的ＨＣＬＶ疫苗株，在长期的使用传代过程中发生了一定程度的遗传变异。In this report, we showed the amplification of the major antigenic protein E1 (gp55) gene of two Chinese hog cholera virus strains,Shimen and 'C'.We accomplished this by designing two set of degenerate primers that were capable of amplifying 5' -terminal half and 3' -terminal half of E1 gene respectively.In addition, the cDNA cloning and nucleotide sequencing of the amplified E1 genes were performed.The amino acid sequence data derived showed that:A.the number and location of cysteine residues in E1 amimo acid sequence of strain'C' and strain Shimen were identical to those of reference strains Alfort,Brescia,ALD and GPE -,revealing that antigenic E1 of HCV had a conserved structure frame; B.the signal sequence at N-terminal and the transmembrane sequence at C terminal were highly conserved,however the antigenic regions located at N-terminal half of E1 were variable,especially at the region of amino acid residues 702-742; C.Comparison of amino acid sequences of E1 between the strain'C' used in our study and the strain'C' used in Rijn's study in the Netherlands showed some amino acid substitutions which scattered along the whole E1 sequence,indicating the strain 'C' might vary genetically to some extents in the course of many years application.

关 键 词：猪瘟病毒 gp55 序列分析 克隆 

分 类 号：S858.285.3[农业科学—临床兽医学]