抗A型产气荚膜梭菌α毒素单链抗体基因的表达  

Expression of ScFv Genes against Alpha toxin of Clostridium perfringens Type A

作　　者：赵宝华[1] 许崇波[2] 冯书章[2] 朱平[2] 

机构地区：[1]河北师范大学生命科学院,河北石家庄050016 [2]解放军军需大学军事兽医研究所,吉林长春130062

出　　处：《中国兽医学报》2000年第4期367-370,共4页Chinese Journal of Veterinary Science

基　　金：全军医药卫生青年基金课题 !( 98Q0 76)

摘　　要：将 2种抗 A型产气荚膜梭菌α毒素单链抗体 ( Sc Fv)基因 Sc Fv-2 E3和 Sc Fv-1 A8克隆至表达载体p UC1 1 9、p HOG2 1和 p ET2 0 b中 ,构建了重组质粒 p UC2 E3和 p UC1 A8、p HOG2 E3和 p HOG1 A8、p ET2 E3和 p ET1 A8后 ,分别转化至受体菌 JM1 0 5、JM1 0 9和 BL2 1 ( DE3 )中 ,得到重组菌株 JM1 0 5( p UC2 E3 )和JM1 0 5( p UC1 A8)、JM1 0 9( p HOG2 E3 )和 JM1 0 9( p HOG1 A8)及 BL2 1 ( DE3 ) ( p ET2 E3 )和 BL2 1 ( DE3 )( p ET1 A8)。 ELISA检测表明 ,经 IPTG诱导后所表达的目的蛋白存在于重组菌株 JM1 0 5( p UC2 E3 )和JM1 0 5( p UC1 A8)及 JM1 0 9( p HOG2 E3 )和 JM1 0 9( p HOG1 A8)的菌培养上清中 ,在重组菌株 BL 2 1 ( DE3 )( p ET2 E3 )和 BL2 1 ( DE3 ) ( p ET1 A8)的胞周质中检测到了 Sc Fv的存在。 SDS-PAGE和薄层扫描分析表明 ,重组菌株 BL2 1 ( DE3 ) ( p ET2 E3 )和 BL2 1 ( DE3 ) ( p ET1 A8)的蛋白表达产物分别占菌体可溶性蛋白的 0 .9%和 0 .7%,其大小约为 2 60 0 0 ,且表达的目的蛋白具有中和磷脂酶 C的活性。The recombinant plasmid pUC2E3 and pUC1A8 , pHOG2E3 and pHOG1A8,pET2E3 and pET1A8 were obtained by the ScFv 2E3 and ScFv 1A8 gene fragments to be isolated and inserted into the expression vectors pUC119, pHOG21 and pET20b. Then the recombinant plasmids were transformated into E.coli for construcion of the recombinant strains.When these strains were induced by IPTG,the ScFv protein could be produced. The expressed ScFv was proved by ELISA and SDS PAGE analysis.in the culture medium of the strains JM105(pUC2E3),JM105(pUC1A8), JM109(pHOG2E3) and JM109(pHOG1A8) and in the supernatant and precipitation of lysised bacteria cell of BL21(DE3)(pET2E3) and BL21(DE3)(pET1A8).The expressed products were about 0.9% and 0.7 % of soluble cellular protein. The expressed ScFv could neutralize phospholipase C.

关 键 词：A型产气荚膜梭菌 Α毒素 单链抗体 基因表达 

分 类 号：S855.12[农业科学—临床兽医学] S852.616.3[农业科学—兽医学]