首页 | 本学科首页   官方微博 | 高级检索  
     检索      

草鱼出血病病毒VP6蛋白的原核表达、纯化及免疫效果
引用本文: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.
作者姓名: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
作者单位:1. 苏州大学基础医学与生物科学学院,江苏苏州,215123
2. 江苏省水生动物疫病预防控制中心,江苏南京210036;南京师范大学生命科学学院,江苏南京210046
3. 江苏省水生动物疫病预防控制中心,江苏南京,210036
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)
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500μg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%。

关 键 词:草鱼出血病病毒  原核表达  纯化  免疫
收稿时间:11/3/2011 3:14:36 PM
修稿时间:12/8/2011 3:40:45 PM

Prokaryotic expression,purification and immune efficacy of VP6 protein of grass carp reovirus
LIU Lin,XU Shi-ying,LI Jing-hui,ZOU Yong,NI Jin-di,YANG Yuan-jie,CAO Guang-li,XUE Ren-yu,CHEN Hui and GONG Cheng-liang.Prokaryotic expression,purification and immune efficacy of VP6 protein of grass carp reovirus[J].Journal of Fisheries of China,2012,36(3):429-435.
Authors:LIU Lin  XU Shi-ying  LI Jing-hui  ZOU Yong  NI Jin-di  YANG Yuan-jie  CAO Guang-li  XUE Ren-yu  CHEN Hui and GONG Cheng-liang
Institution:1(1.School of Biology & Basic Medical Sciences,Soochow University,Suzhou 215123,China; 2.Jiangsu Center for Control and Prevention of Aquatic Animal Infectious Disease,Nanjing 210036,China; 3.College of Life Sciences,Nanjing Normal University,Nanjing 210046,China)
Abstract: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.
Keywords:grass carp reovirus  prokaryotic expression  purification  immunize
本文献已被 CNKI 万方数据 等数据库收录!
点击此处可从《水产学报》浏览原始摘要信息
点击此处可从《水产学报》下载免费的PDF全文
设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号