摘要
为获取具有免疫原性的猪流行性腹泻病毒S蛋白,并能有效研究其免疫原性,从山东省猪流行性腹泻病毒感染的某猪场病料中扩增S蛋白基因具有免疫原性的4个片段(Sa,Sb,Sc和Sd)。将得到的目的片段导入原核表达载体pGEX-6P-1中,转化到BL21(DE3)感受态细胞中进行表达。将表达的蛋白免疫家兔以研究其免疫原性。结果表明,IPTG诱导融合蛋白表达条件的最佳诱导时间为8h,IPTG的最佳终浓度为1.0mmol/L。SDS-PAGE和Western-blot分析表明,四个表达产物的分子质量分别约为61ku(Sa)、46ku(Sb)、41ku(Sc)、43ku(Sd)。经琼脂扩散试验测定,它们免疫后效价分别约为24(Sa)、25(Sb)、22(Sc)、24(Sd)。该研究表明,表达的4段蛋白均具有免疫原性,这为后期亚单位疫苗的研制奠定了基础。
To obtain porcine epidemic diarrhea virus(PEDV)S protein fragments with immunogenicity,four immunogenic fragments(Sa,Sb,Sc,and Sd)of PEDV S gene were amplified and cloned into pGEX-6P-1prokaryotic expression vector,and transformed into Escherichia coli BL21(DE3)competent cells for expression.When rabbits were immunized with the refolding proteins,immunogenicity was detected.In result,the optimized inducible time was 8h,and the optimized inducible concentration of IPTG was 1.0mmol/L.SDS-PAGE and Western-blot analyses showed that Sa was 61ku,Sb 46ku,Sc 41ku and Sd 43ku.Identified by the agar diffusion test,Sa was 24,Sb 25,Sc 22 and Sd 24.The result indicated that the 4expressed proteins were immunogenic,which was useful for development of subunit vaccines against porcine epidemic diarrhea.
出处
《中国兽医科学》
CAS
CSCD
北大核心
2014年第7期752-757,共6页
Chinese Veterinary Science
关键词
猪流行性腹泻病毒
S基因
表达
免疫原性
porcine epidemic diarrhea virus
S gene
expression
immunogenicity
