传染性法氏囊病超强病毒株VP2基因原核表达及其免疫原性检测  

Prokaryotic expression of very virulent infectious bursal disease virus VP2 gene and immunogenic detection of expression product

作　　者：朱彩霞[1] 朱瑞良[1] 刘冠华[1] 翁立雪[1] 马荣德[1] 孙寅剑[1] 

机构地区：[1]山东农业大学动物科技学院山东省动物生物技术与疫病防治重点实验室,山东泰安271018

出　　处：《中国预防兽医学报》2011年第2期154-156,共3页Chinese Journal of Preventive Veterinary Medicine

基　　金：高等学校博士学科点专项科研基金(060434011);山东省科技攻关项目(2008GG10009023);农业部公益性行业科研专项(200803019);山东省博士后科研项目择优资助经费(200603040)

摘　　要：为检测传染性法氏囊病超强毒株(vvIBDV)重组VP2蛋白的免疫原性,本研究利用RT-PCR方法扩增vvIBDV的结构蛋白VP2基因,并将其克隆到表达载体pGEX-4T-3中,构建重组表达质粒pGEX-VP2。将其转化受体菌E.coli BL21(DE3)plysS,经IPTG诱导后,SDS-PAGE电泳和western blot分析表明,表达的重组蛋白约69 ku,并以包涵体形式存在。表达的重组蛋白经纯化后,免疫6周龄BALB/c小鼠制备免疫血清,ELISA分析表明制备的血清效价在1∶5 120以上,表明vvIBDV VP2具有良好的免疫原性,为建立vvIBDV的ELISA检测方法提供了试验依据。To express the VP2 gene of very virulent infectious bursal disease（vvIBDV）,the vvIBDV VP2 gene was amplified by RT-PCR and was inserted into vector pGEX-4T-3 to construct recombinant plasmid pGEX-VP2.The pGEX-VP2 was transformed into E.coli BL21（DE3） plysS and induced with IPTG.The results of SDS-PAGE and western blot indicated that the expressed VP2 protein was about 69 ku and mainly existed in form of inclusion bodies.The anti-VP2 serum was prepared in BALB/c mice immunized with purified VP2 with the titer of 1：5,120,which showed that the express product of VP2 had immunogenic reaction.

关 键 词：传染性法氏囊病超强病毒株 VP2 E.coliBL21(DE3)plysS 表达 

分 类 号：S852.61[农业科学—基础兽医学]