土耳其斯坦东毕吸虫原肌球蛋白基因的原核表达  

Prokaryotic expression of tropomyosin gene of Orientobilharzia turkestanicum

作　　者：王春仁[1] 仇建华[2] 何国声[3] 周庆民[2] 邢继兰[3] 许腊梅[2] 

机构地区：[1]黑龙江八一农垦大学动物科技学院,黑龙江大庆163319 [2]黑龙江省兽医科学研究所,黑龙江富裕161200 [3]中国农业科学院上海家畜寄生虫病研究所,上海200232

出　　处：《中国兽医科学》2006年第3期212-215,共4页Chinese Veterinary Science

基　　金：黑龙江省"十五"科技攻关计划项目(GC01B511-01)

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。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（Ni^2＋） 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.

关 键 词：土耳其斯坦东毕吸虫 原肌球蛋白基因 原核表达 

分 类 号：S852.735[农业科学—基础兽医学] Q786[农业科学—兽医学]