CAJ | 학술논문

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。
목적제비간라간균갑기기단추화신호전도단백다극륭항체。방법장아실보충적중조질립pET22b+/MCP전화대장간균감수태BL21(DE3)，IPTG유도표체His-rhMCP융합단백，장표체적융합단백순화후，면역가토제비항체，간접ELISA법측정항체효개，이제비적균체외막단백（간라간균、담형라간균급유문라간균）채용Western blot감정항체적특이성。결과대장간균성공표체His-rhMCP융합단백，ELISA법검측항체적효개체도1∶32000，Western blot검측결과현시항체가여간라간균균체외막단백급His-rhMCP특이결합。결론제비료고효개、고특이성적rhMCP항체，위진일보연구간라간균적류행병학전정료기출。
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.