肝螺杆菌甲基基团趋化信号转导蛋白多克隆抗体的制备及鉴定.PDFVIP

J South Med Univ, 2013, 33(9): 1295-1298 doi 10.3969/j.issn. 1673-4254.2013.09.08 ·1295 · 基础研究 肝螺杆菌甲基基团趋化信号转导蛋白多克隆抗体的制备及鉴定肝螺杆菌甲基基团趋化信号转导蛋白多克隆抗体的制备及鉴定 1 2 2 2 2 何胜平 ，陈雅华 ，赵瑛瑛 ，王继德 ，白 杨 1 2 南方医科大学南方医院 胸心血管外科，消化内科，广东 广州 510515 摘要：目的制备肝螺杆菌甲基基团趋化信号转导蛋白多克隆抗体。方法 将我室保种的重组质粒pET22b+/MCP 转化大肠杆菌 感受态BL21(DE3) ，IPTG诱导表达His- rhMCP 融合蛋白，将表达的融合蛋白纯化后，免疫家兔制备抗体，间接ELISA 法测定抗 体效价，以制备的菌体外膜蛋白（肝螺杆菌、胆型螺杆菌及幽门螺杆菌）采用Western blot 鉴定抗体的特异性。结果大肠杆菌成 功表达His- rhMCP 融合蛋白，ELISA 法检测抗体的效价达到1∶32000 ，Western blot 检测结果显示抗体可与肝螺杆菌菌体外膜 蛋白及His- rhMCP 特异结合。结论 制备了高效价、高特异性的rhMCP 抗体，为进一步研究肝螺杆菌的流行病学奠定了基础。 关键词：肝螺杆菌；甲基基团趋化信号转导蛋白；抗体制备 Preparation and identification of polyclonal antibody against methyl-accepting chemotaxis signal transduction protein of Helicobacter hepaticus 1 2 2 2 2 HE Shengping , CHEN Yahua , ZHAO Yingying , WANG Jide , BAI Yang 1Department of Cardiovascular Thoracic Surgery, 2Department of Gastroenterology, Nanfang Hospital, Guangzhou 510515, China Abstract: Objective To prepare the polyclonal antibody against methyl-accepting chemotaxis signal transduction protein (MCP) of Helicobacter hepaticus (H.hepaticus). Methods The recombinant plasmid pET22b +/MCP was transformed into E.coli BL2l(DE3) to express the fusion protein His-rhMCP under the induction of IPTG. The fusion protein was purified and the antibody was obtained by immunizing rabbits. The titer of the polyclonal antibody was tested by indirect ELISA, and the specificity of the antibody was identified based on Western blotting using the prepared cell surface proteins (CSPs) of the bacteria. Results The fusion protein was successfully expressed, and the titer of the antibody reached 1 ∶32 000. Western blotting in