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伤寒沙门菌线性质粒pBSSB1的p17基因的生物学功能分析

, PP. 168-173

Keywords: 伤寒沙门菌,线性质粒,pBSSB1,p17

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Abstract:

H:z66阳性伤寒沙门菌含有一特有的介导鞭毛单向相变换的线性质粒pBSSB1,其上第17号基因(p17)编码一未知功能蛋白(P17)。利用λ-Red系统制备p17基因缺陷变异株,测定变异株和野生株的生长曲线,比较其生长差异,同时通过半固体LB培养基进行动力试验比较其动力差异。利用PCR技术扩增获得p17基因,构建其重组表达载体pET28a(+)-p17,通过镍柱纯化目的蛋白P17-His6,纯化后蛋白作为抗原免疫兔子以制备多克隆抗体。结果显示,制备了p17基因缺陷变异株,变异株的生长明显迟缓于野生株(P<0.05),变异株的动力明显减弱。构建了重组表达载体pET28a(+)-p17,纯化获得了高纯度的目的蛋白P17-His6并制得相应的多克隆抗体。

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