| 鸡胚组织膜蛋白与鸡毒霉形体黏附蛋白的结合分析 |
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| 引用本文： | 彭秀丽,高俊伟,胡思顺,胡福利,毕丁仁.鸡胚组织膜蛋白与鸡毒霉形体黏附蛋白的结合分析[J].畜牧兽医学报,2008,39(12). |
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| 作者姓名： | 彭秀丽 高俊伟 胡思顺 胡福利 毕丁仁 |
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| 作者单位： | 1. 华中农业大学动物医学院,武汉,430070;华中农业大学动物遗传育种与繁殖教育部重点实验室,武汉,430070
2. 华中农业大学动物医学院,武汉,430070 |
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| 基金项目： | 湖北省科技攻关重大专项基金 |
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| 摘 要： | 利用pGEX融合蛋白表达系统，将鸡毒霉形体黏附蛋白（pMGA）与GST的编码序列在大肠杆菌BL21中进行融合表达，经GST·BindResin纯化，GST—pMGA的纯度达96％。用蛋白酶Thrombin切掉GST标签，获得纯度为96％的pMGA，经Western—blot鉴定具有良好的免疫学活性。通过差速离心法提取SPF鸡胚的气管、心脏、肝脏、肺脏、肾脏、腺胃、十二指肠、法氏囊、胸腺和脾脏等组织膜蛋白，采用斑点酶联免疫吸附试验（Dot—ELISA）检测各组织膜蛋白与pMGA的结合。结果表明：除脾脏外，其余组织的膜蛋白与pMGA之间存在特异性结合，说明这些组织膜蛋白中存在pMGA的受体蛋白。经SDS-PAGE分析发现，除脾脏外，其余9种组织的膜蛋白中均含有一条相对分子质量约为30ku的主带，受体的相对分子质量可能为30ku，可为深入研究pMGA的受体蛋白提供资料。
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| 关 键 词： | 鸡胚 组织膜蛋白 鸡毒霉形体黏附蛋白 Dot—ELISA |
Binding Research Between the Tissues Membrane of Chick Embryo and pMGA |
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| PENG Xiu-li,GAO Jun-wei,HU Si-shun,HU Fu-li,BI Ding-ren.Binding Research Between the Tissues Membrane of Chick Embryo and pMGA[J].Acta Veterinaria et Zootechnica Sinica,2008,39(12). |
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| Authors: | PENG Xiu-li GAO Jun-wei HU Si-shun HU Fu-li BI Ding-ren |
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| Institution: | PENG Xiu-li1,2,GAO Jun-wei1,HU Si-shun1,HU Fu-li1,BI Ding-ren 1*(1.College of Veterinary Medicine,Huazhong Agricultural University,Wuhan 430070,China2.Key Laboratory of Agricultural Animal Genetics,Breeding , Reproduction of Ministry of Education,China) |
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| Abstract: | The GST-pMGA was expressed in E.coli BL21 with the pGEX fusion expression system.GST-pMGA expressed in supernatant was purified using GST Bind Resin and a plenty of soluble protein was obtained,the final preparation was 96% after purification by SDS-PAGE analysis.GST-pMGA was incubated with thrombin protease overnight,the pMGA was obtained and showed well immunological activity by Western-blot analysis.The tissues membrane protein from trachea,heart,lung,liver,kidney,thymus,bursa of Fabricius,spleen,provent... |
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| Keywords: | Dot-ELISA |
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