文山松毛虫质型多角体病毒(DpwCPV)多角体蛋白基因的cDNA克隆及序列分析  

Cloning and Sequencing of Dendrolimus Punctatus Wenshanensis Cytoplasmic Polyhedrosis Virus (DpwCPV) Polyhedrin Gene

作　　者：王琼[1] 张珈敏[1] 文力[1] 杨娟[1] 胡远扬[1] 

机构地区：[1]武汉大学病毒研究所,湖北武汉430072

出　　处：《武汉大学学报（理学版）》2002年第4期475-480,共6页Journal of Wuhan University:Natural Science Edition

基　　金：国家高技术产业及示范工程项目资助

摘　　要：通过对文山松毛虫质型多角体病毒的增殖 ,纯化 ,获得一株单一类型的质型多角体病毒 提纯的病毒粒子经SDS 酚抽提 ,琼脂糖凝胶电泳分离基因组dsRNA ,低熔点琼脂糖电泳法回收纯化编码多角体蛋白的第十片段S10 S10经DMSO变性 ,逆转录合成cDNA第一条链 ,PCR扩增后 ,克隆在PMD18 T载体上 ,克隆片段全长 76 3bp 起始密码AUG位于第 3～ 5位碱基 ,终止密码UGA位于第 74 7～ 74 9位碱基 推测DpwCPV多角体蛋白基因编码一段由 2 4 9个氨基酸组成的多肽 ,分子质量为 2 84 39.4 4× 10 3 与舞毒蛾质型多角体病毒 (LdCPV)多角体蛋白基因比较 ,两者核苷酸序列的同源性为 97% ,它们各自编码的多角体蛋白的氨基酸同源性为 99% ;与家蚕质型多角体病毒(BmCPV)多角体蛋白基因相比较 ,两者核苷酸序列的同源性为 88% ,它们各自编码的多角体蛋白的氨基酸同源性为 97%Amplify and purify the Dendrolimus punctatus wenshanensis cytoplasmic polyhedrosis virus (DpwCPV) and abstract their nuclears. To syntheses the cDNA of polyhedin gene of DpwCPV, it was denaturalized by DMSO. After RT\|PCR, the amplified cDNA was cloned into the PMD18\|T Vecter. The polyhedrin gene of DpwCPV was found to be 764 nuleotides in length with one reading frame in one strand. It can code a predicted protein of 249 residues( M r of 28 439.44×10 3). Comparison of the nucleotide sequence of the polyhedrin gene of DpwCPV with that of LdCPV and BmCPV showed that nucleotide homology is 97% and 88%, but there is only 1 amino acid difference between DpwCPV and LdCPV and 5 amino acid differences between DpwCPV and BmCPV.

关 键 词：文山松毛虫 质型多角体病毒 多角体蛋白基因 序列分析 

分 类 号：Q785[生物学—分子生物学]