【摘要】 本试验用Ｔｒｉｓ－ＨＣｌ缓冲液（Ｔ）、ＰＢＳ缓冲液（Ｐ）、生理盐水（Ｓ）和２Ｍ尿素ＰＢＳ缓冲液（Ｕ）以热处理的方法提取大肠杆菌Ｋ９９和Ｆ４１菌毛抗原，并用硫酸铵盐析纯化Ｋ９９以及用１Ｎ醋酸滴定和氯化镁沉淀分别纯化Ｆ４１。根据ＳＤＳ－ＰＡＧＥ电泳和蛋白质含量测定结果，四种缓冲液对Ｋ９９的提取量依次是Ｕ＞Ｓ＞Ｐ，而Ｔ检不出Ｋ９９；对Ｆ４１的提取量依次是Ｕ＞Ｔ＞Ｐ＞Ｓ。Ｋ９９纯化后取得了满意的纯化结果，Ｆ４１纯化后大量上样做电泳时仍有微细的污染带出现，而且氯化镁沉淀后造成Ｆ４１严重损失。Ｋ９９和Ｆ４１纯化前后的各个样品经超声波处理、０．５％脱氧胆酸钠处理和未处理的原液与自家因子血清做琼扩试验以及Ｋ９９和Ｆ４１做交叉琼扩试验，均具有良好的特异性，而且以超声波处理后的样品做琼扩试验效果最佳。更多还原

【Abstract】 PiLi antigens K99 and F41 of enterotoxigenic E、coli were txtrated by heat tresatment with Tris HCI(T),PBS (P),0 85% NaCl (S) and 2M urea PSB(V),then K99 antigen was purified by ammonium salfate precipitation,and F41 antigen was purified by titration with TN acetic acid and MgCl 2 precipitation respectively The SDS PAGE and guantity of protein showed that the quantity of K99 antigen in the extracted buffer was V>S>P,but undetectable in buffer T,the quantity of F41 antigen was V>T>P>S The SDS PAGE showed the purification of K99 were satisfied,but fine polluted bands were present in large amounts of F41 purifications and F41 antigen was greatly lost during MgCl 2 Precipitation.Though all of the untreated,ultrasonicates treated and 0 5% deoxycholate treated antigens K99 and F41 of dlifferent purified stages exhibited high specificity in gel double diffusion test with absorbed K99 and F41 antisera and cross gel double diffusion test K99 antigens and F41 antisera,K99 antisera and F41 antigens,the ultra sonicates treated samples txhibited better results更多还原