肠毒素大肠杆菌CS3定居因子抗原和融合肠毒素基因在减毒痢疾杆菌中的共表达  

Simultaneous Expression of CS3 Colonization Factor Antigen and LT-B/ST Fusion Enterotoxin Antigen of Enterotoxigenic Escherichia coli by Attenuated Shigella flexneri 2a

作　　者：刘陶陶[1] 李淑琴[1] 张兆山[1] 郑继平[1] 刘秀丽[1] 罗刚[1] 黄翠芬[1] 

机构地区：[1]北京生物工程研究所,100071

出　　处：《生物化学与生物物理学报》2003年第1期49-54,共6页

基　　金：国家高技术研究发展计划 ( 86 3计划 ) (No.2 0 0 1AA2 15 2 11);全军科研基金重点课题 (No.0 1Z0 2 6 )资助~~

摘　　要：肠毒素和定居因子抗原 (CFAs)是肠毒素大肠杆菌 (ETEC)两种主要的致病因素。有效的ETEC疫苗应包括这两种成分。采用基因重组技术 ,将定居因子CFA/II的共有抗原成分CS3菌毛抗原和LT B/ST融合肠毒素基因克隆至以asd基因为选择标记的重组质粒上 ,与asd基因剔除的弗氏志贺氏菌Fwl0 1构成了宿主 载体平衡致死系统。结果表明 ,在无抗生素条件选择的情况下 ,该重组菌可稳定表达CS3菌毛抗原和LT B/ST融合肠毒素抗原。通过口服和鼻饲方式免疫小鼠 ,可诱生相应的血清IgG抗体 ,同时能够检测到分泌型IgA的产生 ,表明该重组菌可以有效的诱导产生粘膜免疫。Enterotoxigenic Escherichia coli (ETEC) causes watery dehydrating diarrhea in infants in developing countries, and is the most common cause of travelers’ diarrhea. It has been known that the colonazition factor antigens(CFAs) and enterotoxins are important virulence factors of ETEC, and these two kinds of proteins should be included in any effective vaccine against ETEC. In this study, a host-plasmid lethal balancing system was constructed based on asd gene in an avirulent strain of S.flexneri to express CS3 antigens and the fusion LT-B/ST enterotoxins of Escherichia coli. Both of these antigens were expressed steadily in the S. flexneri vector without any antibiotic markers. Antibodies against CS3, LT, ST and LPS of Shigella were detected in sera of mice that were immunized with recombinant bacteria either oragastrically (o.g.) or intranasally (i.n.). SIgA against CS3 and enterotoxins were detected simultaneously in feces of mice. This work is helpful for constructing multivalent recombinant vaccine for prevention of bacterial diarrhea.

关 键 词：CS3菌毛 弗氏志贺氏菌 宿主-载体平衡致死系统 肠毒素大肠杆菌 CS3定居因子 抗原 融合肠毒素 减毒痢疾杆菌 共表达 

分 类 号：Q786[生物学—分子生物学]