| Ara h2三聚体重组蛋白的制备及其低致敏原性鉴定 |
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| 引用本文： | 易海涛,汤慕瑾,刘志刚,闫浩,夏立新. Ara h2三聚体重组蛋白的制备及其低致敏原性鉴定[J]. 免疫学杂志, 2011, 0(10): 840-844 |
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| 作者姓名： | 易海涛 汤慕瑾 刘志刚 闫浩 夏立新 |
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| 作者单位： | 深圳大学过敏反应与免疫学研究所;深圳出入境检验检疫局; |
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| 基金项目： | 国家自然科学基金(30871752); 深圳出入境检验检疫局科技计划项目(SZ2008105); 深圳大学创新团队基金资助项目(200904) |
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| 摘 要： | 目的制备花生主要过敏原Ara h 2三聚体重组蛋白并检测其过敏原性。方法利用分子生物学的方法将3分子的Ara h 2依次串联起来,并将其整合到原核表达载体pET-32a(+),再转化到感受态Origami中;然后利用IPTG诱导其表达;通过Ni2+亲和层析纯化三聚体重组蛋白;Western-blotting和ELISA检测目的蛋白的过敏原性。结果测序结果表明Trimer成功整合到pET-32a(+)上。三聚体重组蛋白纯化后经SDS-PAGE鉴定,蛋白大小与理论值相符。Western-blotting和ELISA结果表明Trimer与重组的Ara h 2(r-Ara h 2)蛋白相比,结合花生过敏病人混合血清中IgE的能力有所降低。结论成功制备花生主要过敏原Ara h 2三聚体重组蛋白,初步的体外实验表明该重组蛋白具有低致敏原的潜能。
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| 关 键 词： | 花生过敏原 Arah2 三聚体 低致敏 |
Preparation and hypoallergenic characterization of Ara h 2 Trimer |
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| YI Haitao,TANG Mujin,LIU Zhigang,YAN Hao,XIA LixinAllergy , Immunology Institute of Shenzhen University,Shenzhen ,Chin. Preparation and hypoallergenic characterization of Ara h 2 Trimer[J]. Immunological Journal, 2011, 0(10): 840-844 |
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| Authors: | YI Haitao TANG Mujin LIU Zhigang YAN Hao XIA LixinAllergy Immunology Institute of Shenzhen University Shenzhen Chin |
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| Affiliation: | YI Haitao,TANG Mujin,LIU Zhigang,YAN Hao,XIA LixinAllergy and Immunology Institute of Shenzhen University,Shenzhen 518060,Chin |
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| Abstract: | This study is aimed to express and purify the peanut major allergen Ara h 2 Trimer and preliminarily characterize the hypoallergenic of purified recombinant trimer protein.Three Ara h 2 molecules were connected and inserted into the expression vector pET-32a(+).Then the vector was transformed into Origami and the protein expression was induced by IPTG.The recombinant trimer protein was purified by Ni2+ chelating affinity chromatography.And the hypoallergenic of Ara h 2 trimer protein was examined by Western... |
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| Keywords: | Peanut allergen Ara h 2 Trimer Hypoallergenic |
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