【摘要】 将pGEM-TM质粒上的土耳其斯坦东毕吸虫原肌球蛋白(TM)基因片段亚克隆至原核表达载体pET28a(+),重组的pET28-TM在大肠埃希氏菌BL21(DE3)中经1 mmol/L IPTG诱导表达出一37.5 ku的融合蛋白。该蛋白经Ni-NTA亲和层析柱纯化,SDS-PAGE检测,出现与目的蛋白大小一致的单一条带。Western-blotting检测结果表明,纯化的蛋白可被自然感染东毕吸虫的山羊血清识别,这为进一步研究东毕吸虫基因工程疫苗奠定了基础。更多还原

【Abstract】 The cloned TM gene fragment from the recombinant plasmid pGEM-TM was subcloned into the expression plasmid pET28a（+）.The recombinant plasmid pET28-TM was transformed into E.coli BL21（DE3） and induced with 1mmol/L of IPTG.The recombinant pET28-TM produced a recombinant protein with an apparent molecular weight of 37.5ku,which was identical to the expected weight.The expressed protein was purified by meta(Ni²⁺) chelation affinity chromatography.The purified protein was proved to be antigenic by Western-blotting analysis using goat serum infected naturally with Orientobilharzia turkestanicum.The results provided foundation for the development of an recombinant vaccine against Orientobilharzia turkestanicum.更多还原