谷胱甘肽S-转移酶和乳酸脱氢酶在亚洲带绦虫囊尾蚴中的表达

摘要

为开发相关疫苗和治疗靶分子,应用免疫组化、双向电泳(2-DE)结合蛋白质印迹(Western-blotting)技术对亚洲带绦虫囊尾蚴谷胱甘肽S-转移酶(GST)和乳酸脱氢酶(LDH)的表达进行了研究.结果表明:亚洲带绦虫囊尾蚴表达GST,囊尾蚴的双向电泳凝胶共检测到204±11个蛋白质斑点,相对分子质量为Mr14400～94000,等电点(pI)为3.0～10.0;Western-blotting分析显示,GST和LDH特异性抗原抗体阳性杂交斑点均为1个,阴性对照均未见阳性杂交斑点;将Western-blotting检测的抗原抗体阳性杂交斑点与原双向电泳凝胶斑点进行比对,均找到对应蛋白斑点,经ImageMaster 2D Platinum 6.0软件分析后初步确定,该蛋白斑点的pI/Mr分别为6.5/25 588和8.2/35 318,与亚洲带绦虫GST和LDH的pI/Mr理论推导值接近.亚洲带绦虫囊尾蚴表达GST和LDH.%Expression of glutathione s-transferase (GST) and lactate dehydrogenase (LDH) in cysticercus of Taenia asiatica was studied by immunohistochemistry method (except LDH) and two dimensional electrophoresis (2-DE) combined with western-blotting to develop relative vaccine and treat target molecule. The results indicated that GST could be expressed in cysticercus of T. asiatica, 204±11 protein spots were detected by 2-DE gel of T. asiatica, its relative molecular mass (Mr) was 14400～94000, and polymorphic loci (PI) was 3.0～10.0. Analysis of western-blotting indicated that there was 1 positive hybridization spot of antibody for specific antigen of both GST and LHD, and there was no spot for negative control. Corresponding protein spot was found in comparison of hybridization spot detected by both western-blotting and 2-DE, and its ratio of PI/Mr was 6. 5/25588, and 8. 2/35318 by analysis of software Image Master 2D Platinum 6.0, which was close to the theoretical prediction value of GST and LHD in T. asiatica. Therefore the study proposed that the GST and LDH could be expressed in cysticercus of T. asiatica express.