Construction of attenuated Salmonella typhimurium Strain expressing Helicobacter pyloriconservative region of adhesin antigen and its immunogenicity
Construction of attenuated Salmonella typhimurium Strain expressing Helicobacter pyloriconservative region of adhesin antigen and its immunogenicity作者机构：PLAInstituteforDigestiveMedicineNanfangHospitaltheFirstMilitaryMedicalUniversityGuangzhou510515GuangdongProvinceChina InstituteofBiotechnologyAcademyofMilitaryMedicalSciencesBeijing100071China
出 版 物：《World Journal of Gastroenterology》 (世界胃肠病学杂志（英文版）)
年 卷 期：2004年第10卷第17期
页 面：2498-2502页
核心收录：
学科分类：1007[医学-药学(可授医学、理学学位)] 100705[医学-微生物与生化药学] 1002[医学-临床医学] 1001[医学-基础医学(可授医学、理学学位)] 100103[医学-病原生物学] 10[医学]
基 金：Supported by the National Natural Science Foundation of China No.30270078
主 题：沙门氏菌属 盲肠疾病 疲劳反应 哈比特属 幽门菌 附着因子 保守治疗 抗原 免疫原性 消化系统
摘 要：AIM: To construct a non-resistant and attenuated Salmonella typhimurium ( S. typhimurium) strain which expresses conservative region of adhesion AB of Helicobacter pylori(Hpylon) and evaluate its ***: The AB gene amplified by PCR was inserted into the expression vector pYA248 containing asd gene and through two transformations introduced into the delta Cya1 delta Crp1 delta Asd attenuated Salmonella typhimurium strain, constructing balanced lethal attenuated Salmonella typhimurium strains X4072 (pYA248-AB). Bridged ELISA method was used to measure the expression of AB antigen in sonicate and culture supematant. According to the method described by Meacock, stability of the recombinant was evaluated. Semi-lethal capacity test was used to evaluate the safety of recombinant. The immunogenicity of recombinant was evaluated with animal ***: The attenuated S. typhimurium X4072 (pYA248-AB) which expresses AB was successfully ***, bridged ELISA assay showed that the content of AB in recombinant X4072 (pYA248- AB) culture supematant was higher than that was in thallus lytic liquor. And after recombinant X4072 (pYA248- AB) was cultured for 100 generations without selection pressure, bhe entire recombinant bacteria selected randomly could grow, and the AB antigen was defected positive by ELISA. The growth curve of the recombinant bacteria showed that the growth states of X4072 (pYA248) and X4072 (pYA248-AB) were basically consistent. The survival rate of C57BL/6 was still 100%, at 30 d after mice taking X4072 (pYA248-AB) 1.0×l0^10 cfu orally. Oral immunization of mice with X4072 (pYA248-AB) induced a specific immune ***: In vitro recombinant plasmid appears to be stable and experiments on animals showed that the recombinant strains were safe and immunogenic in vitro,which providing a new live oral vaccine candidate for protection and care of H pylori infection.
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