摘要
目的利用平衡致死系统构建表达O157:H7 EspA的减毒鼠伤寒沙门氏菌。方法构建表达EspA的重组质粒,再将其转入终宿主菌减毒鼠伤寒沙门氏菌x4550株中构建成口服活疫苗株,用IPTG进行诱导表达,经聚丙烯酰胺凝胶电泳、Westen-blot检测EspA蛋白的表达情况。并观察重组菌体外培养的稳定性。结果利用宿主-载体平衡致死系统构建了表达O157:H7 EspA的重组减毒沙门氏菌,经Tricine-SDS-PAGE电泳出现了1条Mr约21000的蛋白条带,Westen-blot检测能与抗EspA的单克隆抗体发生特异性反应。且在没有选择压力的条件下体外能稳定地繁殖、生长和传代。结论表达O157:H7 EspA的重组减毒沙门氏菌构建成功,为发展口服抗EHEC O157:H7的疫苗奠定了初步基础。
Objective To construct an attenuated Salmonella typhimurium vaccine strain expressing the gene EspA of EHEC O157： H7 with balanced lethal system. Methods The gene of EspA of EHEC O157：H7 was amplified, and then recombined with a vector （asd^＋）. The recombinant was transfected into attenuated Salmonella typhimurium x4550 The expression of EspA was analyzed by Tricine-SDS-PAGE and Western blotting. The stability of vaccine strains was investigated by generation culture in vitro. Results The stable attenuated Salmonella typhmurium vaccine strains expressing O157：H7 EspA was constructed with balanced lethal system. T The expressed products with Mr21000 could react with the antibody of EspA. Conclusion The attenuated Saalmonella typhimurium vaccine strain could probably serve as an oral live vaccine against O157 ：H7 infection.
出处
《免疫学杂志》
CAS
CSCD
北大核心
2007年第2期187-190,共4页
Immunological Journal
