A型产气荚膜梭菌α毒素的基因扩增和提纯鉴定  

AMPLIFICATION AND IDENTIFICATION OF ALPHA TOXIN GENE IN Clostridium perfringens TYPE A

作　　者：于立新[1] 苏佳[1] 赵晓慧[1] 马学恩[1] 么宏强[1] 

机构地区：[1]内蒙古农业大学兽医学院,呼和浩特010018

出　　处：《内蒙古农业大学学报（自然科学版）》2011年第4期34-37,共4页Journal of Inner Mongolia Agricultural University(Natural Science Edition)

基　　金：内蒙古自然科学基金资助项目(31060332)

摘　　要：为了对A型产气荚膜梭菌α毒素快速准确地做出检测,本实验利用厌氧肉肝汤培养A型产气荚膜梭菌,经革兰染色观察以及含铁牛奶培养基观察其生化特性。然后使用饱和度为60﹪的硫酸铵粗提其α毒素蛋白,进行透析纯化并应用SDS-PAGE方法鉴定A型产气荚膜梭菌的α毒素,鉴定之后将纯化的α毒素给小鼠腹腔注射来检测其活性。结果表明,厌氧肉肝汤培养基变浑浊且产生大量气体,含铁牛奶培养基发生爆烈发酵现象,SDS-PAGE检测结果,α毒素分子量为43.6kDa,小白鼠出现梭菌病的典型临床症状。本实验为α毒素的鉴定及大量快速提取提供了一种参考方法,为进一步开展梭菌毒素的致病机理及制备类毒素疫苗奠定重要的实验基础。To detect Clostridium Perfringens type A exactly and quickly,the biochemical characteristics of CVCC38 Clostridium Perfringens type A were determined by culturing in anaerobic meat liver soup,Gram＇s stain and culturing in iron milk medium in this study.Alpha toxin protein was roughly extracted by using ammonium sulfate（saturation for 60%） to dialysis purify and identify it by SDS-PAGE.Bioactivity of the purified alpha toxin was detected by injected into mouse peritoneal cavity.The results showed that anaerobic meat liver soup was turned turbid and produced large amounts of gas;the iron milk medium produced blasting fierce fermentation;alpha toxin molecular weight was 43.6kDa;the mouse appeared clinical symptoms of clostridial disease.This study made important experimental basis for identification and extraction of alpha toxin to explore the pathogenesis mechanism of it and prepare the toxoid vaccine.

关 键 词：产气荚膜梭菌 Α毒素 基因扩增 毒素鉴定 

分 类 号：S852.3[农业科学—基础兽医学]