【摘要】 利用电击转化法将表达绿色荧光蛋白（green fluorescent protein,GFP）的原核表达载体pTurboGFP-B转化到产肠毒素大肠杆菌（enterotoxigenic Escherichia coli,ETEC）F4ac中，获得表达绿色荧光的菌株ETEC F4ac-GFP。聚合酶链式反应（polymerase chain reaction,PCR）结果显示，转化菌株表达绿色荧光蛋白编码基因，且可以在蓝光仪和荧光显微镜下清晰地观察到转化菌株发出绿色荧光；将ETEC F4ac-GFP与IPEC-J2细胞系进行体外粘附试验，在细胞水平验证了转化菌株荧光表达的稳定性与可靠性，且ETEC F4ac导入荧光质粒后不改变其对宿主细胞的胞粘附能力。CCK-8(Cell Counting Kit-8)和实时荧光定量PCR(qRT-PCR)结果表明，ETEC F4ac导入荧光质粒后不改变其对宿主细胞的毒力。本研究为深入探究ETEC F4ac致病机理提供了工具菌株。更多还原

【Abstract】 One strain named as ETEC F4ac-GFP expressing green fluorescent protein(GFP)was obtained through transforming prokaryotic expression vector pTurboGFP-B into enterotoxigenic Escherichia coli(ETEC)F4ac by electrotransformation. It was identified that,by polymerase chain reaction(PCR),the obtained strain not only expressed the gene coding GFP,but also sent out green fluorescence as observed under a blue ray or a fluorescence microscope. The adhesion capacity of ETEC F4ac-GFP to IPEC-J2 cell lines was tested by in vitro adhesion test to verify the stability and reliability of its fluorescent expression at cell level,and the adhesion capacity to host cells was not changed when fluorescent plasmids were input. It was observed that,by cell counting kit-8(CCK-8)and realtime fluorescence quantitative PCR(qRT-PCR),the virulence of ETEC F4ac to host cells was not changed when fluorescent plasmids were input. A strain was thus provided as a tool for further researches on the pathogenesis of ETEC F4ac.更多还原