| 小西葫芦黄花叶病毒外壳蛋白抗体制备 |
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| 引用本文： | 祁伟,韦传宝,杜宇.小西葫芦黄花叶病毒外壳蛋白抗体制备[J].生物学杂志,2010,27(1):35-38. |
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| 作者姓名： | 祁伟 韦传宝 杜宇 |
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| 作者单位： | 1. 安微大学生命科学学院,合肥,230032
2. 皖西学院化学与生命科学系,六安,237012 |
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| 基金项目： | 安徽省教育厅青年项目基金 |
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| 摘 要： | 根据已报道的小西葫芦黄花叶病毒（ZYMV）序列设计特异引物，扩增ZYMV的全长外壳蛋白（CP）基因，插入原核表达载体pSBET后在大肠杆菌B121（DE3）plysS中诱导表达。通过12％SDS—PAGE和5％～20％梯度SDS—PAGE二次制备电泳纯化诱导产物，免疫小鼠，获得经过Western blot分析为特异的抗CP血清。硫酸铵沉淀法与ProteinA—Red Sepharose亲和层析相结合提取IgG，获得效价达1：4800的一抗，对西瓜和甜瓜田间样品的间接ELISA检测表明，ZYMV在田间普遍发生，研究制备的IgG可用于ZYMV检测。
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| 关 键 词： | 小西葫芦黄花叶病毒 原核表达 抗血清制备 IgG提取 间接ELISA |
Prokaryotic expression of ZYMV CP gene and preparation of the antibody against CP |
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| QI Wei,WEI Chuan-bao,DU Yu.Prokaryotic expression of ZYMV CP gene and preparation of the antibody against CP[J].Journal of Biology,2010,27(1):35-38. |
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| Authors: | QI Wei WEI Chuan-bao DU Yu |
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| Institution: | QI Wei1,WEI Chuan-bao2,DU Yu2(1.School of Life Science,Anhui University,Hefei 230032,2.Department of Chemistry , Biology,West Anhui University,Liuan 237012,China) |
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| Abstract: | Specific primer was designed to amplify ZYMV complete coat protein(CP) gene.The CP gene was then inserted into pSBET vector and expressed in Escherichia coli BL21(DE3) plys S strain.The object protein was purified by 12%SDS-PAGE firstly and subsequently 5%~20% gradient SDS-PAGE.The antiserum against the CP was prepared from mice and its specificity was confirmed by Western blot analysis.IgG against ZYMV-CP was purified by ammonium sulfate sedimentation and Protein A-Red Sepharose affinity chromatography.The... |
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| Keywords: | Zucchini yellow mosaic virus prokaryotie expression antiserum preparation IgG purification indirect ELISA |
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