南方医科大学学报

南方医科大学学报 ›› 2013, Vol. 33 ›› Issue (09): 1295-.

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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

何胜平，陈雅华，赵瑛瑛，王继德，白杨   

  • 出版日期:2013-09-20 发布日期:2013-09-20

Preparation and identification of polyclonal antibody against methyl-accepting chemotaxis signal transduction protein of Helicobacter hepaticus

  • Online:2013-09-20 Published:2013-09-20

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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。

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 indicated that the antibody could specifically bind to CSPs and His-rhMCP. Conclusion The antibody with a high titer
and specificity was prepared to facilitate further study of the pathogenicity and epidemiology of H.hepaticus in human.