利用T4噬菌体展示猪瘟病毒E2抗原  

CSFV E2 Antigen Display on the Surface of Bacteriophage T4

作　　者：吴健敏[1] 任兆钧[2] 余兴龙[1] 涂长春[1] 张念祖 

机构地区：[1]解放军军需大学军事兽医研究所 [2]美国马里兰州大学医学院生物化学及分子生物研究中心,巴尔的摩美国212011503 [3]云南省热带亚热带动物病毒病开放实验室,昆明650024

出　　处：《中国生物工程杂志》2004年第11期61-64,共4页China Biotechnology

基　　金：国家"973"计划资助项目 (G19990 1190 3 )

摘　　要：利用重组PCR技术将猪瘟病毒 (CSFV )E2蛋白主要抗原编码区基因 (mE2 )与T4噬菌体SOC基因融合 ,构建了大小为 643bp的SOC mE2融合基因 ,再将其插入携带T4溶菌酶基因 (e)和(denV)基因的T4重组载体 (PRH) ,构建了重组载体pRsmE2。通过重组载体与缺失突变型T4发生同源重组 ,可将SOC mE2融合基因整合入T4的基因组中 ,并成功地将大小约 2 1 5aaSOC mE2融合蛋白展示于T4噬菌体衣壳表面。经Westernblot、胶体金免疫电镜等免疫学检测证实 ,展示于T4表面的mE2融合蛋白具有CSFV免疫学活性。The T4 phage display is a newly developed system for displaying of exogenous peptides or proteins domains by fusion expression on the T4 surface. In this system the displayed proteins maintain their relatively independent conformation and biological activities. This process is initiated by fusing a foreign gene to the T4 surface capsid accessory protein gene soc (small outer capsid protein). The protective antigenic mE2(encoding 132aa most antigenic peptide at N termianl of E2) gene of classical swine fever virus (CSFV), was fused in frame to 3′ end of soc in display vetor. The soc/ mE2 fusion genes were then integrated into the soc site of the genome of soc-deleted T4 phage mutant by homologous recombination leading to the display of the mE2 protein on the surface of T4 phage. In western blot the correct bands corresponding to the molecular weights of fused soc/mE2 proteins were visualized. The immunogold staining electromicroscopy showed the display proteins were correctly located on the surface of recombination of T4 phage particles. Inoculation of mice with recombinant phage T4.soc.mE2 showed a significant immune response as indicated in ELISA using intact CSF virus or E.coli expressed E2 protein as antigens. The detectable antibody level persisted for 56 days of a complete immunization trial. ;

关 键 词：重组载体 融合基因 抗原编码区 融合蛋白 噬菌体展示 携带 免疫学检测 猪瘟病毒 CSFV 利用 

分 类 号：Q78[生物学—分子生物学] S852.651[农业科学—基础兽医学]