摘要
为了比较VP2和NS1两种蛋白的免疫原性,选择免疫原性较好的蛋白进行亚单位疫苗制备。本试验分别扩增了水貂细小病毒(mink enteritis virus,MEV)NS1与VP2基因,连接pET-32a表达载体并进行表达,对表达产物进行SDS-PAGE及Western blotting分析。以His-Bind亲和层析柱纯化目的蛋白,将纯化后的蛋白免疫小鼠,分析目的蛋白的免疫原性。经SDS-PAGE与Western blotting鉴定,表明NS1与VP2蛋白大小分别为83和67ku,且均具有生物学活性;免疫小鼠后,目的蛋白NS1和VP2均可诱导小鼠产生抗MEV特异性抗体,且VP2蛋白诱导小鼠产生的抗体滴度要高于NS1蛋白。与NS1蛋白比较,VP2蛋白更适合亚单位疫苗的制备。
In order to develop subunit vaccine of mink enteritis virus,the immunogenicity of mink parvovirus protein NS1 and VP2had been evaluated.Two pairs of primers were designed,and the full-length NS1 and VP2genes had been amplificated,and then prokaryotic expression vector pET-32a-NS1,PET-32a-VP2 were constructed.After the analysis of SDS-PAGE and Western blotting,target proteins had been purified by His-Bind affinity chromatography.The immunogenicity of purified protein NS1 and VP2 were evaluated by serum ELISA testing,after inoculated BALB/c mouse.The results showed that the molecular mass of NS1 and VP2protein were 83and67 ku by SDS-PAGF and Western blotting;Although both target protein NS1 and VP2had the ability to induce BALB/c mouse to produce anti-MEV specific antibodies,the level of antibodies induced by the protein VP2 was higher than protein NS1.Mink parvovirus protein VP2 was more suitable for the development of subunit vaccine.
出处
《中国畜牧兽医》
CAS
北大核心
2016年第6期1630-1634,共5页
China Animal Husbandry & Veterinary Medicine
基金
吉林省科技发展计划项目(20130206026NY)
关键词
水貂细小病毒
原核表达
重组蛋白
mink parvovirus
prokaryotic expression
recombinant protein
