【摘要】 目的 :研究金黄色葡萄球菌 (金葡菌 )毒素的调控机制 ,构建pET_trap表达载体 ,在大肠杆菌中表达金葡菌毒素调控的重要分子TRAP蛋白。方法 :从金葡菌中提取基因组DNA ,用特异引物钓取trap基因。通过亚克隆的方法构建了pET_trap表达载体 ,在大肠杆菌中诱导表达 ,经亲和柱层析分离纯化。制备纯化蛋白的多抗血清 ,利用Western印迹检测多抗与天然蛋白的反应。结果 :目的基因在大肠杆菌中获得高表达 ,超声破碎后 ,表达产物主要存在于上清中 ,制备的多抗血清可以与天然蛋白发生特异性反应。结论 :通过原核表达获得了天然的TRAP蛋白 ,为研究金葡菌毒素调控机制及防治金葡菌感染奠定了基础更多还原

【Abstract】 Objective: To study the regulation of virulence factors secreted by Staphylococcus aureus , and to acquire and express trap gene in E.coli . Methods:The trap gene was acquired by PCR amplification from S.aureus genome with specific primers.The expression vector pET_trap was constructed and expressed in E.coli . The TRAP protein was purified by affinity chromatography. The polyclonal antibodies against TRAP protein were prepared and tested by Western Blot. Results: The TRAP protein was highly expressed in E.coli in soluble state. The prepared antibody can specifically recognize the natural TRAP protein. Conclusion:The acquirement of purified TRAP protein will lay a foundation for the study of the regulation of virulence factors in S.aureus .更多还原