SARS-CoV核衣壳蛋白的表达与免疫研究  

Expression of the Gene Encoding the N Protein of SARS Virus in E. coli and Its Usd in the Immunization of Hourse

作　　者：王瑞琳[1] 金宁一[1] 赵博[2] 金洪涛[1] 贾雷立[1] 金明兰[1] 

机构地区：[1]军事医学科学院军事兽医研究所,吉林长春130062 [2]吉林大学生命科学学院,吉林长春130023

出　　处：《中国病毒学》2005年第6期582-585,共4页Virologica Sinica

基　　金：吉林省科技厅重大项目(20030413)

摘　　要：依据GenBank中SARS基因组序列,采用人工合成的方法合成编码SARS病毒N蛋白的全基因(1296bp)序 列,再与设计的CTL特异性表位基因(195bp)重组后,克隆到pET-28a(+)质粒中,重组质粒转化到大肠杆菌BL21 (DE3)中诱导表达。利用SARS患者恢复期阳性血清,鉴定表达蛋白。进一步纯化后免疫马,并用ELISA方法检 测血清抗体效价。结果显示SDS-PAGE表明所表达的蛋白相对分子质量约为55000 Da,与预计大小相符;Western blot显示表达的蛋白具有良好的抗原性和特异性。对表达蛋白形成的包涵体进行洗涤,得到的蛋白纯度可达到 70．1％。切胶纯化免疫马后,获得的抗SARS抗体滴度达1:2560。为亚单位疫苗研制和精制抗SARS抗体免疫制 剂提供基础。The gene encoding the N protein of SARS virus was cloned and expressed in E. coli. The recombinant protein was identified by anti-SARS positive sera obtained from recovered SARS patients. The recombinant N protein （55 kDa） reacted specifically with anti-SARS positive sera. The recombinant N protein was purified and used to immunize horses, and the antibody was detected. The recombinant protein of SARS virus displayed specific reaction with SARS antibody. The results showed that the horses immunized with this recombinant protein produced higher antibody titers （1：2560）. Our results provide information on the immunization against the SARS virus.

关 键 词：严重急性呼吸道综合症(SARS) 核衣壳蛋白 N 表达 抗体 IGG 

分 类 号：R392[医药卫生—免疫学]