小麦丛矮病毒M蛋白基因的序列测定、表达和鉴定  

Cloning, Expression and Identification of Matrix Protein Gene of Wheat Rosette Stunt Virus (WRSV)

作　　者：叶永钧[1] 龚祖埙[1] 

机构地区：[1]中国科学院上海生物化学研究所

出　　处：《生物化学与生物物理学报》1998年第5期520-524,共5页

基　　金：国家自然科学基金

摘　　要：从小麦丛矮病毒（ｗｈｅａｔｒｏｓｅｔｅｓｔｕｎｔｖｉｒｕｓ，ＷＲＳＶ）基因文库含磷蛋白ＮＳ基因的质粒中，经亚克隆、测序，得到ＮＳ蛋白基因的下游序列，其中含有一读框４５３ｎｔ、编码一分子量约１７ｋＤ蛋白。用ＰＣＲ方法获得该读框全长片段并克隆到表达载体ｐＧＥＸ－３Ｘ，在大肠杆菌ＢＬ２１（ＤＥ３）菌株中以ＩＰＴＧ诱导表达，产物由谷胱甘肽Ｓ－转移酶（ＧＳＴ）亲和层析柱纯化后，用蛋白质印迹分析鉴定，结果表明，该读框为编码病毒基质蛋白Ｍ的基因。cDNA fragment located downstream of the WRSV NS protein gene was sequenced. Following an 80 nt untranslated region at the 5′terminus, there is a 453 nt open reading frame(ORF) encoding a 17 kD protein, which was allowed to be expressed in E.coli using bacterial expression vector pGEX 3X and produced a 43 kD fusion protein. The result of Western blot showed that the fusion protein was able to react strongly with antibody raised against the purified WRSV particles. According to the similarities between the gene organizations of VSV and WRSV and between the molecular weights of deduced and expressed proteins, as well as the viral structural M protein, and to the result of Western blot, the 453 nt ORF was identified as WRSV M protein gene.

关 键 词：小麦丛矮病毒 基质蛋白 核酸顺序 表达 免疫印迹 

分 类 号：S435.121.9[农业科学—农业昆虫与害虫防治] Q78[农业科学—植物保护]