致病性鸡大肠杆菌type1菌毛fimC基因的克隆与序列测定

Cloning and sequence analysis of type1 pili fimC gene from avian pathogenic Escherichia coli (APEC)

根据国外发表的人致病性大肠杆菌fimC基因序列,在其保守区设计了带有BamHⅠ/HindⅢ酶切位点的一对引物,应用PCR技术以鸡致病性大肠杆菌O2基因组DNA为模板扩增到一个片段,大小约为700bp,将扩增产物克隆到pMD18＿T载体上,并转化于大肠杆菌TG1宿主菌,经酶切和筛选,得到阳性重组质粒。通过对阳性重组质粒核酸序列测定,确定此DNA片段为鸡大肠杆菌fimC基因。

According to the published fimC gene sequence of type1 pili from human Escherichia coli, a pair of primers with two enzyme digest sites of BamHⅠ and HindⅢ was designed and synthesized,which localize conservative regions of fimC gene. One fragment was amplified by polymers chain reaction(PCR)with genomic DNA extracted from Escherichia coli strain O2 as template.length is 700 bp,Amplicions were cloned into pMD18-T plasmid vector with E.coli TG1 as host .The recombinant plasmid was obtained by screen and enzyme digest,after sequence analysis of this recombinant plasmid,the result showed that the DNA fragment was fimC gene from avian E.coli.