传染性法氏囊病病毒结构蛋白VP3基因的克隆与原核表达  

Molecular cloning and prokaryotic expression of structural protein VP3 gene of IBDV

作　　者：彭志伟[1] 王笑梅[1] 高宏雷[1] 付朝阳[1] 张厚双[1] 包晓玮[2] 冉多良[2] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所,黑龙江哈尔滨150001 [2]新疆农业大学,新疆乌鲁木齐830052

出　　处：《中国预防兽医学报》2006年第2期124-127,共4页Chinese Journal of Preventive Veterinary Medicine

摘　　要：利用PCR技术扩增出IBDV的结构蛋白VP3基因,将其克隆到表达载体pPROEX-HTa中,获得重组质粒命名为VP3/PA,经PCR、酶切和序列分析鉴定表明,插入的位置、大小和读码框均正确。SDS-PAGE检测表明,经重组质粒VP3/PA转化、诱导的受体菌E.coliDH5α能表达结构蛋白VP3基因,约33Ku,表达量达到了菌体总蛋白的33.9%,经Westernblot等检测表明,诱导表达的抗原蛋白能与vvIBDV阳性血清发生特异性反应。这为IBDV血清学诊断方法的建立打下了基础。By PCR, a structural protein gene VP3 of infectious bursal disease virus （IBDV） was determined from segment A of IBDV .The gene was cloned into expressing vector pPROEX-HTa.The recombinant plasmid named VP3/PA was constructed.The VP3/PA was used to transform into E.coli DH5α.The results of SDS-PAGE and Western blot indicated that the VP3 protein was expressed in high level （33.9 % ） and the recombinant fusion protein,which was about 33 Ku,had immunological reactive activity.This study lay on foundation for the development of the diagnosis methods in serology for IBDV.

关 键 词：传染性法氏囊病病毒 结构蛋白VP3基因 克隆与原核表达 

分 类 号：S852.659.5[农业科学—基础兽医学] Q785[农业科学—兽医学]