绿脓杆菌PA-SD-1株外毒素AⅢ区基因的克隆及序列分析

Clonging and Sequencing of the Exotoxin A DomainⅢ Gene Fragment of Pseudomonas aeruginosa

利用PCR技术 ,以绿脓杆菌羊分离株 (暂命名为PA_SD_1株 )的染色体DNA为模板 ,成功扩增出了该菌的主要致病基因 (绿脓杆菌外毒素A)Ⅲ区基因。将该PCR扩增片段与pGEMR_TEasy载体连接 ,获得了含有目的片段的重组质粒 (命名为PA_SD_ETA)。序列测定结果表明。该菌株与其他已报道的绿脓杆菌菌株有较高的同源性 ,而局部出现单个碱基的点突变。

Using polymerase chain reaction technique,DomainⅢ gene of Exotoxin A was amplified from the chromosomal DNA of one field isolates of sheep pseudomonas aeruginosa.the PCR amplified gene was linked with pGEM R-T Easy Vector,then transformed into E.coli strain DH5α,the recombinant plasmid contained 0.658kb fragment was obtained.Sequece analysis showed that the isolate from sheep shared high identity with the other isolates from human.