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土耳其斯坦东毕吸虫原肌球蛋白基因的原核表达
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王春仁1，仇建华2，何国声3，周庆民2，邢继兰3，许腊梅2
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（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）
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摘要：
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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1mmol/L IPTG诱导表达出一37.5ku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。 |
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关键词：
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土耳其斯坦东毕吸虫；原肌球蛋白基因；原核表达 |
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中图分类号：S 852.735：Q786 文献标识码：
A 文章编号：1673-4696(2006)03-0212-04 |
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Prokaryotic expression of tropomyosin gene of
Orientobilharzia turkestanicum |
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WANG Chun-ren1, QIU
Jian-hua2, HE Guo-sheng3, ZHOU
Qing-min2, XING Ji-lan3, XU
La-mei2 |
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(1. College of Animal Science and Technology, Heilongjiang August First Land Reclamation University, Daqing
163319， China; 2. Heilongjiang Provincial Institute of Veterinary Science, Fuyu 161200，China; 3.Shanghai Institute
of Domestic Animal Parasitology, Chinese Academy of Agricultural Sciences, Shanghai 200232， China) |
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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 1mmol/L of IPTG. The recombinant
pET28-TM produced a recombinant protein with an apparent molecular weight of 37.5ku,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. |
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Key words: Orientobilharzia turkestanicum; tropomyosin gene; prokaryotic expression |
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