【摘要】 为研究毒素源性大肠杆菌(ETEC)菌毛蛋白分子装配机理,从K88adETEC中PCR扩增出K88菌毛蛋白小亚基基因片段faeC,然后将其克隆到大肠杆菌表达载体pET30a(+)中,获得重组质粒pET30C,并将该质粒转入大肠杆菌表达菌株BL21(DE3)中;重组菌株经IPTG诱导后,经过SDS PAGE分析表明该菌株可以表达FaeC蛋白(16.9kDa),且重组蛋白主要以包涵体形式存在,约占菌体蛋白的30%。用分离纯化的重组FaeC蛋白免疫小鼠,获得FaeC的鼠抗血清,经免疫学分析表明,利用此重组蛋白制备的抗血清与FaeC重组蛋白及K88adETEC的菌毛蛋白都能发生特异性抗原抗体反应。更多还原

【Abstract】 To study the assembly mechanism of ETEC (enterotoxigenic E. coli) pilin molecules, the gene segment of faeC, encoding a subunit of K88 pilin, was amplified by PCR from ETEC, and then the segment was cloned into E. coli expression vector pET-30a(+), resulting in the recombinant plasmid pET30C, which was subsequently transferred into E. coli strain BL21(DE 3). After the recombinant strain was induced by IPTG, the SDS-PAGE analysis showed that the strain could express FaeC protein (16.9 kDa), and the recombinant protein occupied about 30% of total bacterial proteins with inclusion bodies. The recombinant FaeC protein was purified, and used to immunize a mouse to obtain its polyclonal antibodies. The immunological analysis indicated that this antiserum had a specific antigen-antibody reaction with the FaeC recombinant protein and the pilin of K88 ad ETEC.更多还原