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 cterminal gene of alphatoxin(cpa408) was amplified from the strain NCTC64609 ofClostridium perfringens (Type A) by PCR and then was cloned into a clone vector pMD18T.By PCR and EcoR 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 Cterminal fragment of alphatoxin gene of the foreigners over 99%,which proved that the cloned gene was the Cterminal fragment of alphatoxin gene.
出处
《生命科学研究》
CAS
CSCD
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)
关键词
产气荚膜梭菌
Α毒素
基因
克隆
结构分析
Clostridium perfringens Type A
alpha-toxin
c-terminal gene
cloning
nucleotide sequencing
