Preparation and toxicity analysis of recombinant Clostridium perfringens alpha toxin
10.7644/j.issn.1674-9960.2025.05.004
- VernacularTitle:重组产气荚膜梭菌α毒素的制备及毒性研究
- Author:
Pinnan ZHAO
1
;
Xiang GAO
;
Longlong LUO
Author Information
1. 军事科学院军事医学研究院,国家安全特需药品全国重点实验室,北京 100850
- Keywords:
Clostridium perfringens alpha toxin;
prokaryotic expression;
protein purification;
cytotoxicity;
hemolytic activity
- From:
Military Medical Sciences
2025;49(5):349-355
- CountryChina
- Language:Chinese
-
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.
