摘要
用聚合酶链式反应(PCR)技术,从金黄色葡萄球菌中扩增出769bp的编码SPA细胞壁跨膜区的编码序列。将扩增的片断克隆进pEZZ 18载体BamHI和SalI位点之间。重组质粒转化大肠杆菌HB101后,根据酶切分析筛选到含有目的基因的重组质粒pZSPAX。序列测定结果表明,该重组质粒中的插入序列与已发表的SPA X是一致的。通过水肿病毒素B亚单位(SLT IIeB)基因(slt IIeB)证实质粒pZSPAX能作为在细菌表面表达目的基因的载体。
The coding gene sequence of the cell wall's fixed region of SPA(621bp,named SPA-X)is amplified from staphylococcus aurous from animal.Then the PCR products of SPA-X are linked and cloned into prokaryotic excrete expression vector pEZZ-18 and resulting recombinants are called pZSPAX.The SLT-IIe subunit B gene(slt-IIeB)is selected to be a tested gene and cloned into pZSPAX to confirm whether pZSPAX could be a surface display vector.With the technology colloidGold staining,SDS-PAGE and western-blot,it is shown that the recombinant pZSPAX can surface display the gene of SLT-IIe subunit B.
