产气荚膜梭菌α毒素基因的克隆及结构分析  

Cloning and Sequencing of Alpha-toxin Gene of Clostridium perfringens Type A

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

机构地区：[1]军事医学科学院微生物与流行病研究所,中国北京100071

出　　处：《生命科学研究》2003年第2期144-146,共3页Life Science Research

基　　金：全军医药卫生科研基金资助项目(01MB059)

摘　　要：应用PCR技术,从产气荚膜梭菌菌株NCTC64609中,扩增出A型产气荚膜梭菌α毒素基因C端片段(cpa408),并将其克隆至pMD18 T载体中.经转化,α互补蓝白菌落选择培养,提取质粒,进行PCR和EcoRI、PstI双酶切鉴定,筛选出阳性重组克隆.经核苷酸序列分析证实,cpa408基因阅读开放框架由372个核苷酸组成,编码124个氨基酸.经计算机分析,cpa408基因序列与国外文献报道的A型产气荚膜梭菌α毒素基因C端片段同源性达99%以上,表明所克隆的基因即为α毒素基因C端片段.The cterminal gene of alphatoxin(cpa408) was amplified from the strain NCTC64609 ofClostridium perfringens (Type A) by PCR and then was cloned into a clone vector pMD18T.By PCR　and EcoR I/Pst I digestion,the gene was identified.After being sequenced,the cpa408 gene was proved consisting of 372 bp enconding 124 amino acid residues.After being analyzed by computer,the gene was known to be consistent with the published Cterminal fragment of alphatoxin gene of the foreigners over 99%,which proved that the cloned gene was the Cterminal fragment of alphatoxin gene.

关 键 词：产气荚膜梭菌 Α毒素 基因 克隆 结构分析 

分 类 号：Q933[生物学—微生物学] Q754