鸡致病性E.coli O_(24)基因工程包涵体疫苗的制备及免疫原性  

Preparation and Immunity of Genetic Engineering Vaccines from Avian Pathogenic Escherichia coli O_(24)

作　　者：水小溪[1] 李寒鸣[1] 蔡乐[1] 赵宝华[1] 张天兵 

机构地区：[1]河北师范大学生命科学学院,河北石家庄050016 [2]华北制药集团华胜有限公司,河北石家庄052160

出　　处：《河北师范大学学报（自然科学版）》2010年第1期103-107,共5页Journal of Hebei Normal University：Natural Science

基　　金：河北省科技攻关项目(012201130)

摘　　要：利用已构建的基因工程菌株BL21/pET-28a-pilA和BL21/pET-28a-ompC,经IPTG诱导,SDS-PAGE分析分别可见表达的20ku和40.9ku的特异条带,确定蛋白以包涵体的形式存在.Western blot结果表明,表达的Ⅰ型菌毛蛋白(pilA基因编码)和外膜蛋白(ompC基因编码)可与抗体发生特异性结合,说明其具有良好的反应原性.将表达特异蛋白的重组菌株BL21/pET-28a-pilA和BL21/pET-28a-ompC分别制成基因工程包涵体疫苗,免疫小鼠后,具有很好的保护能力.ELISA结果显示,免疫的小鼠体内已产生相应的抗体,效价为1∶400,说明这2种疫苗有很好的免疫原性.表明这2株基因工程菌株有望作为鸡致病性大肠杆菌基因工程疫苗的候选生产菌株.Recombinant strains BL21/pET-28a-pilA and BL21/pET-28a-ompC were induced by IPTG.The specific 20 ku protein and 40.9 ku were detected by SDS-PAGE.Immunogenicity of the type Ⅰ fimbraie（expressing by pilA） and the out membrane protein（expressing by ompC） was confirmed by Western blot.The two recombinant strains BL21/pET-28a-pilA and BL21/pET-28a-ompC expressing type Ⅰ fimbraie and out membrane protein separately were transformed into genetic engineering vaccine.The protective immune response was proved after the mice were immunized with the two vaccines.The titer of antibody in the mice is 1∶400,which was detected by indirect ELISA.The results showed that the two engineering vaccines BL21/pET-28a-pilA and BL21/pET-28a-ompC could be as candidates to provide protective immune response against AEPC infection.

关 键 词：pilA OMPC 基因工程包涵体疫苗 免疫原性 

分 类 号：Q789[生物学—分子生物学]