高价铁肠杆菌素受体蛋白FepA表面抗原决定簇基因的克隆及序列同源性分析

用PCR技术获取高价铁肠杆菌素受体蛋白FepA表面抗原决定簇基因,分析其在大肠杆菌种内的序列同源性。以提取的大肠杆菌基因组DNA为模板,设计一对基因特异引物,用PCR扩增的方法获得目的基因。将PCR扩增后得到的片段纯化并克隆至pMD19-T Simple Vector载体,用菌落PCR及酶切鉴定阳性菌落,对阳性菌落进行测序,并与Gene Bank公布的大肠杆菌区段进行序列同源性分析。扩增出598bp的含有受体蛋白FepA表面抗原决定簇基因的目的片段,经测序分析其种内的序列同源性为95%～99%,为今后制备针对该区段的抗体奠定了基础。

Clone and sequence homology analysis of surface epitope gene in ferric enterobactin receptor protein FepA

The antigen region of the fepA gene of receptor protein fepA was obtained with PCR technology and the sequence homology of it among E.coli was analyzed.With a pair of specific primers,the DNA fragment was amplified by PCR from E.cofi genomic DNA.The PCR amplicons were purified and cloned into pMD19-T Simple Vector.After being identified by PCR amplification and restriction digestion,the positive clones were sequenced and the sequences were biasted against those of fepA gene in E.coli publicated in GenBank.The target gene 598 bp was successfully amplified and the homology were 95% -99%.which was prepared for antibody making.