甜菜坏死黄脉病毒外壳蛋白基因的表达及其表达产物抗血清的制备  

EXPRESSION OF BEET NECROTIC YELLOW VEIN VIRUS COAT PROTEIN GENE AND PREPARATION OF ANTISERUM FROM ITS EXPRESSION PRODUCT

作　　者：姚华建[1] 于嘉林[1] 金元[1] 刘仪[1] 

机构地区：[1]北京农业大学农业生物技术国家重点实验室

出　　处：《病毒学报》1994年第1期39-43,共5页Chinese Journal of Virology

摘　　要：将甜菜坏死黄脉病毒外壳蛋白基因克隆到ｐＢＶ２２０质粒上，构建成在大肠杆菌中经温度诱导表达的载体ｐＢＶＳ４。ＳＤＳ－ＰＡＧＥ电泳和Ｗｅｓｔｅｒｍ印迹结果表明，ｐＢＶＳ４在大肠杆菌ＤＨ５α中通过４２℃诱导后，特异地表达２１ｋＤ的甜菜坏死黄脉病毒外壳蛋白。经光密度扫描估测，其表达量占大肠杆菌全蛋白的１５．２％。分离纯化经温度诱导表达的甜菜坏死黄脉病毒外壳蛋白作为抗原，免疫家兔制备出了特异性强的ＢＮＹＶＶ抗血清。采用微量沉淀法测定其效价为１：１０２４。he temperature-inducible expression vector pBVS4 containing beet necrotic yellow vein vi-rus (BNYVV)coat protein (CP)gene was constructed by cloning the BNYVVCP gene intoplasmid pBV220 and transferred into E.coli DH5α.Expression of the specific protein was achieved by temperature induction.The results of SDS-PAGE and Western blot show thatthe expression product which accumulates 15.2% of the total cellular protein estimated by Shimadzu CS-910 scanning is 21KD BNYVV coat protein.The CP subunit was separated from sodium dodecyl sulfate-polyacrylamide gels and injected into rabbits to prepare antiserum.The titer of antiserum is 1:1024 detected by miniprecipitation.

关 键 词：甜菜 坏死黄脉病毒 外壳蛋白基因 

分 类 号：S432.41[农业科学—植物病理学]