摘要
应用PCR技术,从A型产气荚膜梭菌菌株NCTC64609中扩增出A型产气荚膜梭菌α毒素全基因(cpa1229基因)并将其克隆至pMD18-T载体中。经转化、IPTG/X-gal选择培养,提取质粒,PCR和EcoRI/PstI双酶切鉴定,筛选阳性重组克隆。经核苷酸序列分析证实,cpa1229基因阅读开放框架由1194bp组成。经GenBank检索对照分析,cpa1229基因序列与国外文献报道者同源性达98.3%,表明本实验所克隆的cpa1229基因即为A型产气荚膜梭菌α毒素基因。
The complete gene ofα-toxin (cpa1229gene)was amplified from the strain NCTC64609of Clostridium perfringens type A by PCR,and then was cloned into a clone vector pMD18-T.By PCR and Eco RI /PstI digestion,the gene was identified.After being sequenced,the cpa1229gene was known to be consist of1194bp enconding398amino acid residues.After being analyzed by computer,the gene was known to be consistent with the publishedα-toxin gene of the foreigners up to98.3%.
出处
《生物技术通讯》
CAS
2003年第3期180-181,共2页
Letters in Biotechnology
