迟钝爱德华氏菌鞭毛蛋白的提取与分析  

Purification and analysis of Edwardsiella Tarda flagellin

作　　者：张晓佩[1] 龚晖[2] 陈如敬[2] 杨金先[2] 林天龙[2] 

机构地区：[1]福建师范大学生命科学学院,福建福州350108 [2]福建省农业科学院生物技术研究所,福建福州350003

出　　处：《福建农业学报》2008年第1期35-38,共4页Fujian Journal of Agricultural Sciences

基　　金：农业部科技专项基金(nyhyzx07-043);福建省自然科学基金(2007J0057);福建省科技厅重大专项基金(2004NZ03-2)

摘　　要：采用不同的方法提取迟钝爱德华氏菌鞭毛蛋白(Edwardsiella Tardaflagellin,ETF),选出合适的方法提取ETF,同时制备了ETF的鼠抗血清和迟钝爱德华氏菌菌株ETY全菌的鼠抗血清,通过ELISA法和Western blottig法分析了ETF的抗原性和免疫原性。试验结果表明,酸化高速离心法可获得高纯度分子量约为44 kDa的ETF,该蛋白可被爱德华氏菌的全菌鼠抗血清识别,同时由ETF制备的鼠抗血清除了能特异识别该蛋白之外,也可识别爱德华菌菌体裂解产物中44 kDa的蛋白,证实爱德华氏菌鞭毛蛋白具有抗原性和免疫原性,可作为亚单位疫苗的候选抗原。Methods for isolating Edwardsiella Tarda flagellin （ETF） from E. Tarda were studied . One method was selected. Mouse anti-ETF serum and anti-ETY serum were prepared. The antigenicity and immunogenicity of ETF were analyzed by means of ELISA and Western blotting. The results showed that the method, i. e. , acidification and high-speed centrifuge, could obtain the highly purified ETF, which had a molecular weight about 44kD. The protein was recognized by the mouse anti-ETY and anti-ETF serum. In addition, the mouse anti-ETF serum could recognize the 44kD protein separated from ETY. This proves that ETF possesses antigenicity and immunogenicity. And, therefore, ETF could conceivably be a candidate for the preparation of the subunit vaccine against edwardsiellosis in Japonica eel .

关 键 词：鞭毛蛋白 迟钝爱德华氏菌 提取 抗血清 

分 类 号：Q936[生物学—微生物学]