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.
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
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)
