| 人抗凝血酶Ⅲ(AT-Ⅲ)在毕赤酵母中可溶性表达 |
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| 引用本文： | 于群 宋丽雅 贺敏 卜凤荣 孟志云 孙文种 章金刚. 人抗凝血酶Ⅲ(AT-Ⅲ)在毕赤酵母中可溶性表达[J]. 中国生物制品学杂志, 2004, 17(5): 277-279 |
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| 作者姓名： | 于群 宋丽雅 贺敏 卜凤荣 孟志云 孙文种 章金刚 |
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| 作者单位： | 北京输血研究所,北京100850 |
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| 摘 要： | 目的 研究重组AT Ⅲ基因在巴斯德毕赤酵母菌中进行表达。方法 将携带AT Ⅲ基因的表达载体电转染GS115细胞 ,用G4 18筛选抗性克隆 ,用SDS PAGE及WesternBlot检测表达产物的纯度及特异性 ;用生化法检测AT Ⅲ的表达量 ;用凝血酶空斑法检测表达产物活性。结果 表达产物能与抗AT Ⅲ单克隆抗体结合 ,并具有天然AT Ⅲ的生物活性。结论 已在毕赤酵母菌中成功地表达了AT Ⅲ。
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| 关 键 词： | 抗凝血酶Ⅲ 毕赤酵母 基因表达 |
| 修稿时间： | 2004-01-14 |
Expression of Soluble Recombinant Human Antithrombin Ⅲ (AT-Ⅲ ) in P.pastoris |
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| YU Qun,SONG Li-ya,HE Min,et al. Expression of Soluble Recombinant Human Antithrombin Ⅲ (AT-Ⅲ ) in P.pastoris[J]. Chinese Journal of Bilogicals, 2004, 17(5): 277-279 |
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| Authors: | YU Qun SONG Li-ya HE Min et al |
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| Abstract: | Objective To express human antithrombin Ⅲ(AT-Ⅲ) in P.pastoris.Methods Transfect the eukaryotic expression vector incorporated with AT-Ⅲ gene into P.pastoris GS115 by electroporation, select the recombinants by growth in G418 medium and identify the expressed protein by SDS-PAGE and Western blot.Determine the expression level and activity of AT-Ⅲ by biochemical method and thrombin plaque formation test respectively.Results The expressed protein bound to McAb against AT-Ⅲ and showed biological activity of natural AT-Ⅲ.Conclusion AT-Ⅲ was successfully expressed in P.pastoris. |
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| Keywords: | Antithrombin Ⅲ P.pastoris Gene expression |
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