产气荚膜梭菌贵州分离株α-毒素基因的克隆与鉴定

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

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α 毒素基因的克隆。

The α-toxin genomic DNA containing a specific fragment 242 bp 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.