重组A型产气荚膜梭菌α毒素C端片段的分离纯化  

Isolation and purification of recombinant α-toxin c-terminal domain of Clostridium perfringens type A

作　　者：林明辉[1] 荫俊[1] 邢洪光[1] 王慧[1] 侯晓军[1] 宋伟[1] 

机构地区：[1]军事医学科学院微生物流行病研究所,北京100071

出　　处：《生物技术通讯》2004年第1期26-28,共3页Letters in Biotechnology

摘　　要：押在构建出高表达、高活性的A型产气荚膜梭菌α毒素保护性抗原工程菌株pBV/cpa408的基础上,对表达产物进行了分离纯化研究。诱导表达菌液离心后,超声破碎沉淀细菌,上清用80%饱和硫酸铵沉淀,沉淀蛋白经透析,上凝胶过滤层析柱分离纯化,得到纯度达95%以上的表达目的蛋白,经SDS-PAGE测定,相对分子质量为15.5×103,序列与文献报道的相符。Isolation and purification of recombinant c-terminal domain of Clostridium perfringens type A is studied on the basis of the construction of high expression engineering bacterial pBV/cpa408.After being induced by42℃,the bacterial was centrifuged and lysised by supersonic.The upper fluid was pipeted by80%(NH 4 ) 2 SO 4 and dialysised with pH7.4PBS.Then the expressed protein is isolated and purified through the column of gel filtration chromatography.Analysed by SDS-PAGE,the molecular weight of the expressed protein is about15000Da and its purity is over95%.

关 键 词：C端片段 纯化 Α毒素 分离 A型产气荚膜梭菌 

分 类 号：Q936[生物学—微生物学]