番鸭细小病毒VP2基因在毕赤酵母中的表达及表达产物的鉴定  

Expression of DNA encoding VP2 of MDPV in a pichia pastoris and checking of the expressed products

作　　者：娄华[1] 白挨泉[1] 顾万军[1] 陆英杰[1] 贺东升[2] 刘福安[2] 

机构地区：[1]佛山科技学院生命科学院,广东南海528231 [2]华南农业大学动物医学系,广东广州510642

出　　处：《中国预防兽医学报》2004年第3期202-206,共5页Chinese Journal of Preventive Veterinary Medicine

摘　　要：通过DNA重组技术,将克隆到的MDPV＿Q和MDPV＿26的VP2基因片段,经两种限制性核酸内切酶SacII和Kp nI的酶切,将酶切产物再克隆到表达载体pPICZαA中,得到表达重组质粒pPICZαA＿M＿QVP2和pPICZαA＿M＿26VP2。将这两种表达重组质粒分别转化到毕赤酵母(PichiaPastorisX＿33)中进行表达,经甲醇诱导和SDS＿PAGE分析结果表明,MDPV＿QVP2和MDPV＿26VP2基因片段的表达产物大小约为73kD,与理论预计相符。此外,本研究还意外地发现,重组质粒转化到酵母菌进行表达,在表达VP2结构蛋白的同时,还表达了一条大小约为60kD的蛋白条带。Two recombinant plasmids pMD18-T-M-Q VP2 and pMD18-T-M-26 VP2 were digested with KpnI and SacII . The VP2 inserts were sub-cloned into the expression vector pPICZαA, and recombinant expression plasmids pPICZαA-M-Q VP2 and pPICZαA-M-26 VP2 were obtained.The two recombinant expression plasmids were used to transform into the Pichia Pastoris X-33 and the inserted target genes were expressed.The expressed products were analyzed and checked by SDS-PAGE and Western-blot. Western-blot analysis showed one band of approximately equal intensity that corresponded to the expressed proteins of approximately 73 kD, conforming to theoretical estimate. In addition, an unexpected result was found in that. Western-blot analysis showed two bands;One band was the expected VP2 band, the other was approximatey 60 kD

关 键 词：番鸭细小病毒 毕赤酵母X-33 表达 VP2基因 

分 类 号：S852.659.2[农业科学—基础兽医学]