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产气荚膜梭菌贵州分离株α毒素基因的克隆与鉴定

虞天德1， 王开功2， 王松林3， 孟燕3， 方国升3

(1. 贵州省兽医防疫站, 贵州 贵阳 550008； 2. 贵州大学 动物医学系， 贵州 贵阳 550025;
3. 贵阳市南明区动物检疫所， 贵州 贵阳 550002)

摘要： 提取了A型产气荚膜梭菌贵州分离株的染色体DNA，经聚合酶链反应(PCR)扩增并回收获得了1条242bp的特异性片段，将该基因片段克隆到质粒载体pUC19中，并转化至大肠埃希氏菌JM109中；经氨苄青霉素(Amp)抗药性和显色反应筛选及重组质粒酶切分析、PCR鉴定和Southernblotting检测，表明该重组质粒是产气荚膜梭菌〖WTBX〗α 毒素基因的克隆。

关键词： 产气荚膜梭菌； α 毒素基因； PCR; 克隆； 鉴定

中图分类号：S 852.616.3  文献标识码： A  文章编号：1000-6419(2004)04-0014-04

Cloning and identification of α toxin gene of Clostridiumperfringens Guizhou strain

YU Tiande1, WANG Kaigong2, WANG Songlin3， MENG Yan3, FANG Guosheng3
（1. Guizhou Province Veterinary and Quarantine Station, Guiyang 550000, China；
 2. Department of Veterinary Medicine, College of Animal Science, Guizhou University, Guiyang 550025,China; 3.Nanming District Animal Quarantine Station of Guiyang City ,Guiyang 550002,China)

Abstract: The α toxin genomic DNA containing a specific fragment 242bp in length was extracted from the chromosomal DNA of Clostridium perfringens isolated in Guizhou Province and was amplified by PCR. After the fragment was cloned into pUC19 vector, it was expressed in recombinant strain JM 109. The recombinant plasmid was identified by restriction endonuclease analysis, PCR and Southernblotting. The result showed that the recombinant plasmid is a clone of α toxin gene of Clostridium perfringens .

Key words: Clostridium perfringens ; α toxin gene; PCR; clone; identification

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