禽白血病病毒p27基因在原核细胞的表达及生物学特性的初步分析  

Experession of Avian Leukosis Virus p27 Gene in E.coli and Analysis of Its Biological Characteristics

作　　者：韩静[1] 陈晨[1] 曹红[1] 陈福勇[1] 

机构地区：[1]中国农业大学动物医学物,北京100094

出　　处：《病毒学报》2005年第4期293-297,共5页Chinese Journal of Virology

基　　金：农业部"948"项目资助(编号972081)

摘　　要：将禽白血病病毒(ALV)的p27基因克隆入表达性载体pET28a,在大肠杆菌以HisTag融合蛋白的形式获得了高效表达。以表达产物免疫家兔,制备了抗ALVp27的多克隆抗体,经亲和纯化后,用此抗体建立了对ALV抗原的双抗体夹心法ELISA,并对疑似病料进行了实验室诊断。检测结果与IDEXX的禽白血病抗原检测试剂盒符合率达到98.6%,证明表达产物保留了天然p27蛋白的相关抗原性。交叉试验和群特异性试验证明,此方法具有良好的特异性,并且可以检测出A、BJ、亚群的禽白血病病毒p27抗原。用此ALV抗体和所建立的ELISA方法成功地进行了禽白血病病毒抗原的实验室诊断。The sequence of avian leukosis virus p27 was inserted into pET28a,it expressed in the form of His Tag fusion protein in E.Coli.at high level.The rabbit anti-p27 polyclonal antibody was produced in rabbits immunized with the fusion protein.Using affinity chromatography-purified rabbit anti-p27 polyclonal antibody,we established ALV antigen capture-ELISA.This ELISA coincided with IDEXX ALV antigen capture-ELISA kit up to 98.6%.This result showedthat the expressed fusion protein retained some antigenicities of the nature protein.Cross-activity test and group specificity test justified that this method has good specificity and can successfully detect p27 antigen of ALV ABJ subgroups.Rabbit anti-p27 polyclonal antibody can be successfully used in laboratory diagnosis of ALV by ELISA.

关 键 词：禽白血病病毒 P27基因 生物学特性 初步分析 原核细胞 ELISA方法 实验室诊断 双抗体夹心法 表达产物 多克隆抗体 检测试剂盒 特异性试验 ALV 大肠杆菌 高效表达 融合蛋白 亲和纯化 检测结果 交叉试验 病毒抗原 克隆人 His 

分 类 号：S852.65[农业科学—基础兽医学] TV641.43[农业科学—兽医学]