文章摘要
刘林,徐诗英,李婧慧,邹勇,倪金俤,杨鸢劼,曹广力,薛仁宇,陈辉,贡成良.草鱼出血病病毒VP6蛋白的原核表达、纯化及免疫效果[J].水产学报,2012,36(3):429~435
草鱼出血病病毒VP6蛋白的原核表达、纯化及免疫效果
Prokaryotic expression,purification and immune efficacy of VP6 protein of grass carp reovirus
投稿时间:2011-11-03  修订日期:2011-12-08
DOI:10.3724/SP.J.1231.27803
中文关键词: 草鱼出血病病毒  原核表达  纯化  免疫
英文关键词: grass carp reovirus  prokaryotic expression  purification  immunize
基金项目:苏州市自然科学基金(SYN201106);江苏省科技支撑计划项目(BE2009387);江苏省农业科技自主创新资金项目(SCX(11)2165)
作者单位
刘林 苏州大学基础医学与生物科学学院 
徐诗英 江苏省水生动物疫病预防控制中心 
李婧慧 江苏省水生动物疫病预防控制中心 
邹勇 江苏省水生动物疫病预防控制中心 
倪金俤 江苏省水生动物疫病预防控制中心 
杨鸢劼 江苏省水生动物疫病预防控制中心 
曹广力 苏州大学基础医学与生物科学学院 
薛仁宇 苏州大学基础医学与生物科学学院 
陈辉 江苏省水生动物疫病预防控制中心 
贡成良 苏州大学基础医学与生物科学学院 
摘要点击次数: 2014
全文下载次数: 1598
中文摘要:
      为了探讨草鱼出血病病毒(GCRV)VP6亚单位疫苗对草鱼出血病的免疫保护作用,将vp6基因克隆进原核表达载体 pET-28a(+),构建了重组质粒pET28a(+)-VP6。SDS-PAGE和Western-blotting显示,重组VP6蛋白的分子量约为43 ku,主要以包涵体形式存在,重组蛋白占菌体总蛋白的20%左右;Ni2+柱纯化后的重组蛋白,以每尾500 μg肌肉注射免疫草鱼(14~20 cm,60~120 g),并在第14、21、28、49、70天通过间接凝集反应检测抗体水平,结果显示,免疫注射后第14天可检测到鱼体产生的特异性抗体,第21天达到最高峰,第70天仍可检测到抗体的存在;免疫注射第21天人工接种GCRV,免疫后的草鱼对出血病的保护力达100%。
英文摘要:
      In order to study the immuno-protective efficacy of GCRV-VP6 subunit vaccine,the recombinant expression vector pET28a(+)-VP6 was constructed by cloning vp6 gene of GCRV into the prokaryotic expression plasmid pET-28a(+).The results of SDS-PAGE and Western-blotting showed that the molecular mass of the recombinant VP6 protein was approximately 43 ku.The recombinant protein reached 20% of the total bacterial proteins,and was mainly in the form of insoluble bodies.The grass carp(60-120 g in body weight and 14-20 cm in body length)was immunized with 500 μg purified VP6 protein by Ni2+ affinity chromatography and antibody titers in the blood of fish were determined by means of indirect agglutination reaction on the 14th,21st,28th,49th and 70th days post-vaccination.The results showed that the specific antibody could be detected on the 14th day,reached peak on the 21st day and could be still detected on the 70th day after immunization.The fish in vaccinated group and control group were challenged with GCRV on 21st days after vaccine delivery and the relative survival rate in the vaccined group was 100%.These results provided important academic foundation for research and development of GCRV genetic vaccine.
HTML   查看全文   下载PDF阅读器
关闭

手机扫一扫看