| 猪瘟兔化弱毒E2蛋白A／D区的高效表达和蛋白纯化及其应用 |
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| 引用本文： | 苏小运,缪德年,许立华,陈德胜,陈溥言.猪瘟兔化弱毒E2蛋白A／D区的高效表达和蛋白纯化及其应用[J].Virologica Sinica,2003,18(2):164-168. |
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| 作者姓名： | 苏小运 缪德年 许立华 陈德胜 陈溥言 |
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| 作者单位： | [1]南京农业大学农业部动物疫病诊断与免疫重点开放实验室，江苏南京，210095 [2]南京农业大学农业部动物疫病诊断与免疫重点开放实验室，江苏南京，210095;上海农科院畜牧兽医研究所，上海，201106 [3]南京农业大学农业部动物疫病诊断与免疫重点开放实验室，江苏南京，210095;宁夏大学农学院动物科学系，宁夏银川，750105 |
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| 基金项目： | 国家863高技术发展计划资助项目(2001AA249012) |
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| 摘 要： | 本试验对表达猪瘟E2囊膜糖蛋白的原核表达载体pET—E2的表达条件进行了优化，在此基础上，对表达的蛋白质进行了纯化。在6mol/L盐酸胍存在的条件下，将包涵体溶解在Tris-HCl中，并直接用于His Band亲和层析。收集过柱产物并透析，在2％SDS条件下，对所得的重组蛋白质进行了定量。重组蛋白质被用来包被96孔ELISA板并应用于免疫猪瘟抗体水平的测定。
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| 关 键 词： | 猪瘟兔化弱毒 E2蛋白A／D区 原核表达 E2糖蛋白 EIISA 血清抗体 |
Expression and purification of HCLV E2 protein A, D antigenic domains and its preliminary application |
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| SU Xiao-yun,MIAO De-nian,XU Li-hua,CHEN De-sheng,CHEN Pu-yan.Expression and purification of HCLV E2 protein A, D antigenic domains and its preliminary application[J].中国病毒学(英文版),2003,18(2):164-168. |
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| Authors: | SU Xiao-yun MIAO De-nian XU Li-hua CHEN De-sheng CHEN Pu-yan |
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| Institution: | SU Xiao-yun1,MIAO De-nian1,2,XU Li-hua1,3,CHEN De-sheng1,CHEN Pu-yan1** |
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| Abstract: | In this experiment the conditions for the expressing of HCLV E2 glycoprotein A and D antigenic domains in E coli were optimized. On this basis, the expressed protein was purified by dissolving the inclusion body in Tris-HCI with 6 mol/L guandinine and passed throh a His Bind affinity chromatography. The product was collected and the concentrated recombinant protein was quentified. The recombinant protein was then used to coat 96-well ELISA plates and applied to detect the titers of the serums from swine immunized with HCLV vaccine. |
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| Keywords: | Hog cholera lapinized virus (HCLV) E2 glycoprotein Prokaryotic expressing ELISA Preliminary application |
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