副溶血弧菌VPA1405多克隆抗体制备及应用

发布时间：2018-01-08 23:26

本文关键词：副溶血弧菌VPA1405多克隆抗体制备及应用　出处：《军事医学》2013年02期 　论文类型：期刊论文

【摘要】：目的建立副溶血弧菌(Vibrio parahaemolyticus,VP)的Western印迹方法。方法将VPA1405基因克隆到pET-28a(+)载体,转入大肠杆菌BL21中进行不可溶性表达,在变性条件下纯化得到VPA1405蛋白,制备的包涵体溶液直接免疫兔得到多克隆抗体,进而利用Western印迹检测VPA1405蛋白在野生株(WT)和opaR基因敲除株(ΔopaR)中表达水平的差异。结果成功表达纯化了6个与生物膜相关基因的蛋白;Western印迹结果显示OpaR调控子对VPA1405基因的表达呈正调控,这与表型实验结果相符。结论成功建立了VP的Western印迹实验平台,该平台可用于后续VP生物膜相关基因的调控研究。
[Abstract]:Objective to establish the Vibrio parahaemolyticus (Vibrio parahaemolyticus, VP) Western blot. Methods VPA1405 gene was cloned into pET-28a (+) vector into not soluble expression in Escherichia coli BL21, under denaturing conditions and purified VPA1405 protein by direct solution of the inclusion body prepared rabbit polyclonal antibody was obtained, and then using VPA1405 protein Western was analyzed in the wild strain (WT) and opaR gene knockout strain (opaR) expression level differences. Results the successful expression of the 6 genes associated with biofilm protein purification; Western blot showed that the expression of the positive regulation of VPA1405 gene OpaR promoter, which is consistent with the experimental results. Conclusion the phenotype successfully established Western blot VP platform, the platform can be used to study the regulation of subsequent VP biofilm related genes.

【作者单位】：重庆医科大学公共卫生与管理学院;军事医学科学院微生物流行病研究所病原微生物生物安全国家重点实验室;
【基金】：国家自然科学基金资助项目(31170127,30871370) 国家973计划资助项目(2009CB522604)
【分类号】：R378
【正文快照】： 副溶血弧菌(Vibrio parahaemolyticus,VP)是一种革兰阴性菌,广泛存在于海水、海底沉积物以及鱼、虾、贝、蟹等海洋生物中,是一种重要的食源性致病菌[1]。人们食用被VP污染的食物后,常引起以恶心、呕吐、腹痛、腹泻、发热等为主要症状的急性胃肠炎,其主要特征为水样便,且常混

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