军事医学2025，Vol.49Issue(5)：349-355,7.DOI:10.7644/j.issn.1674-9960.2025.05.004
重组产气荚膜梭菌α毒素的制备及毒性研究
Preparation and toxicity analysis of recombinant Clostridium perfringens alpha toxin
摘要
Abstract
Objective To efficiently produce Clostridium perfringens alpha toxin(CPA)by using a prokaryotic expression system and systematically analyze its biological activity.Methods The CPA gene fragment fused with His-Tag sequence was cloned into a prokaryotic expression vector pTIG-Trx and transformed into E.coli TransB(DE3).The IPTG was used to induce the expression of the target protein,and the protein was obtained by using Ni-column affinity purification.The cytotoxic effect of recombinant CPA protein on 293T,LS174T and SW480 cells,hemolytic effect on human and mouse red blood cells,and lethal effect on mice were further evaluated.Results The recombinant CPA protein with a relative molecular weight of about 45×103 and a purity of more than 90％was successfully obtained.It had significant toxicity to 293T,LS174T and SW480 cells and induced hemolytic reactions in human and mouse red blood cells at specific concentrations.Low dose of CPA protein could cause rapid death in mice in a short time.Conclusion This study successfully obtained the high purity CPA protein with good biological activity in vitro and in vivo,which laid a foundation for further study of the pathogenic mechanism of CPA and its potential application.关键词
产气荚膜梭菌α毒素/原核表达/蛋白纯化/细胞毒性/溶血性Key words
Clostridium perfringens alpha toxin/prokaryotic expression/protein purification/cytotoxicity/hemolytic activity分类
基础医学引用本文复制引用
赵品楠,高翔,罗龙龙..重组产气荚膜梭菌α毒素的制备及毒性研究[J].军事医学,2025,49(5):349-355,7.基金项目
国家自然科学基金项目(82204262) （82204262）
北京市科技新星人才项目(Z211100002121020) （Z211100002121020）
北京市科技新星交叉课题(20220484216) （20220484216）
