福建农业学报2026，Vol.41Issue(4)：421-428,8.DOI:10.19303/j.issn.1008-0384.2026.04.003
金黄色葡萄球菌α-溶血素的原核表达及其活性测定
Prokaryotic Expression and Activity of α-Hemolys in Staphylococcus aureus
摘要
Abstract
[Objective]A prokaryotic expression system of α-Hemolysin(HLα),one of the key virulence factors of Staphylococcus aureus,was constructed to prepare purified recombinant HLα protein(rHLα)with native biological activity,providing important research materials for HLα-targeted drug screening.[Methods]The recombinant plasmid pET30a-hlα was transformed into Escherichia coli BL21(DE3).After expression induction,the inclusion bodies were denatured,purified,and refolded.Hemolytic activity,ability to cause cell membrane damage,and activating effect on host membrane receptor,a disintegrin and metalloprotease 10(ADAM10),of the recombinant protein were evaluated by hemolysis assays,lactate dehydrogenase(LDH)release assays,and western blotting.[Results]The pET30a-hlα recombinant expression system was successfully constructed,and the target protein with a molecular weight of approximately 43 kDa obtained.The rHLα was verified to possess significant hemolytic activity,be able to effectively damage cell membranes at concentrations≥6.3 μg·mL-1,and be capable of inhibiting cellular viability at concentrations≥50 μg·mL-1.In addition,it significantly activated the ADAM10 expression with the toxic effect intensified at increased concentrations.[Conclusion]An efficient and stable rHLα expression and purification system was successfully established that made the recombinant protein with intact biological activity available for further study on the pathogenic mechanism of HLα and development of effective drugs against the bovine mastitis infection.关键词
金黄色葡萄球菌/α-溶血素/膜损伤/ADAM10Key words
Staphylococcus aureus/α-hemolysin/membrane damage/ADAM10分类
农业科技引用本文复制引用
肖冰,薛喜博,毛慧霞,张银花,孙英健..金黄色葡萄球菌α-溶血素的原核表达及其活性测定[J].福建农业学报,2026,41(4):421-428,8.基金项目
北京市自然科学基金项目(6212003) （6212003）
北京农学院科技创新"火花行动"支持计划项目(BUA-HHXD2024004) （BUA-HHXD2024004）
北京农学院大学生创新创业训练计划项目(202510020002X) （202510020002X）
