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伤寒沙门氏菌鞭毛蛋白的分子克隆与表达:伤寒沙门氏菌52 kDa特异性抗原的特性分析

Molecular cloning and expression of Salmonella typhi flagellin: characterization of 52 kDa specific antigen of S. typhi.

作者信息

Sukosol T, Sarasombath S, Mongkolsuk S, Songsivilai S, Chaiyaroj S, Pongsunk S, Ekpo P

机构信息

Department of Immunology, Faculty of Medicine Siriraj Hospital, Bangkok, Thailand.

出版信息

Asian Pac J Allergy Immunol. 1993 Jun;11(1):57-69.

PMID:8216561
Abstract

We previously reported monoclonal antibodies (MAbs) specific to S. typhi 52 kDa antigen which do not cross react with related protein antigens from 11 bacteria causing enteric fever and enteric fever-like illness. Using the combination of these specific MAbs and recombinant DNA technology, expression plasmids containing the antigen gene producing substantial amount of the S. typhi protein antigen have been established. Plasmid pSKM-T7 containing the specific 52 kDa antigen gene was cloned and the antigen expressed was detectable by immunoblotting using specific mAbs. The complete nucleotide sequence of this gene was compared with other bacterial sequences and found to be highly homologous with the flagellin gene H1-d of S. muenchen except in the hypervariable region in the central portion. The specific 52 kDa antigen of S. typhi detected by our MAbs is thus a flagellin.

摘要

我们之前报道过针对伤寒沙门氏菌52 kDa抗原的单克隆抗体(MAbs),这些抗体不会与引起肠热症和肠热症样疾病的11种细菌的相关蛋白抗原发生交叉反应。利用这些特异性单克隆抗体与重组DNA技术相结合,已构建出含有能产生大量伤寒沙门氏菌蛋白抗原的抗原基因的表达质粒。克隆了含有特异性52 kDa抗原基因的质粒pSKM-T7,通过使用特异性单克隆抗体进行免疫印迹可检测到表达的抗原。将该基因的完整核苷酸序列与其他细菌序列进行比较,发现除了中部的高变区外,它与慕尼黑沙门氏菌的鞭毛蛋白基因H1-d高度同源。因此,我们的单克隆抗体所检测到的伤寒沙门氏菌特异性52 kDa抗原是一种鞭毛蛋白。

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