【摘要】 目的:克隆表达伤寒沙门菌高渗诱导基因osmY并制备多克隆抗体。方法:通过PCR技术从伤寒沙门菌基因组DNA中获得伤寒沙门菌高渗诱导基因osmY,再将此基因片段克隆到表达载体pET22b(+)上,并转入大肠埃希菌JM109进行表达;Ni柱纯化后的OsmY蛋白作为抗原免疫家兔,并获得多克隆抗体。结果:成功制备伤寒沙门菌高渗诱导基因osmY的蛋白表达菌株,并获得了纯化的OsmY蛋白,成功制备兔抗OsmY的多克隆抗体。结论:成功克隆表达了伤寒沙门菌高渗诱导基因osmY,并获得了多克隆抗体,有助于今后研究OsmY在伤寒沙门菌高渗应激中的作用。更多还原

【Abstract】 Objective: To clone and express osmotically inducible gene osmY of Salmonella enterica serovar Typhi(S.Typhi),and prepare the anti-OsmY polyclonal antibody.Methods: The DNA fragment of osmotically inducible gene osmY from S.Typhi GIFU 10007 was cloned using PCR.The sequence encoding the protein OsmY was cloned into the pET22b(+) vector and expressed in E.coli JM109.The fusion protein OsmY-His6 was purified by Ni affinity chromatography and was used as antigen to prepare polyclonal antibody.Results: The protein expression strain of osmotically inducible gene osmY was prepared successfully and was highly expressed in E.coli JM109.The anti-OsmY polyclonal antibody was prepared successfully. Conclusion: This study was contributed to research on the role of OsmY in hyperosmotic stress of S.Typhi.更多还原