| 人酸性成纤维细胞生长因子基因在大肠杆菌中的高效表达及产物纯化、鉴定 |
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| 引用本文： | 李兴德,屠重棋,彭红卫,袁淑兰,吴凤锷. 人酸性成纤维细胞生长因子基因在大肠杆菌中的高效表达及产物纯化、鉴定[J]. 四川大学学报(医学版), 1999, 30(3): 249-252 |
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| 作者姓名： | 李兴德 屠重棋 彭红卫 袁淑兰 吴凤锷 |
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| 作者单位： | 1. 成都倍菲生物工程有限公司,成都,610041
2. 华西医科大学附属第一医院 |
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| 摘 要： | 用 Ｈｉｎｄ Ⅲ和 Ｅｃｏ ＲⅠ双酶酶切经改造的 Ｐ Ｕ Ｃａ Ｆ Ｇ Ｆ质粒，获得了人酸性成纤维细胞生长因子（ｈａ Ｆ Ｇ Ｆ）基因，将该基因片段插入表达载体ｐｋｋ２２３３，筛选得到了重组质粒ｐｋｋｈａ Ｆ Ｇ Ｆ。加 Ｉ Ｐ Ｔ Ｇ 诱导该质粒转化的大肠杆菌 Ｊ Ｍ １０９ 表达，ｈａ Ｆ Ｇ Ｆ在发酵液中的表达水平约８０ｍ ｇ／ Ｌ。用肝素亲和层析的方法，获得了电泳纯ｈａ Ｆ Ｇ Ｆ样品。纯化过程中ｈａ Ｆ Ｇ Ｆ活力回收率为６５％ 。纯化ｈａ Ｆ Ｇ Ｆ样品的比活性为 Ｅ Ｄ５０ ４．６ｎｇ／ｍ ｌ，理化及生物活性分析与天然ｈａ Ｆ Ｇ Ｆ对照品完全一致。
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| 关 键 词： | 人酸性成纤维细胞生长因子 基因克隆和表达 分离纯化 鉴定 |
High-level Expression of Human Acidic Fibroblast Growth Factor in E. coli and Its Purification |
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| Li Xingde,Tu Chongqi,Peng Hongwei,Yuan Shulan,Wu Feng'e. High-level Expression of Human Acidic Fibroblast Growth Factor in E. coli and Its Purification[J]. Journal of Sichuan University. Medical science edition, 1999, 30(3): 249-252 |
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| Authors: | Li Xingde Tu Chongqi Peng Hongwei Yuan Shulan Wu Feng'e |
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| Abstract: | s A reconstructed human acidic fibroblast growth factor(haFGF)cDNA was cloned into the expression vector pkk223 3,and the expression in Escherichia coli. JM109 was induced by IPTG induction; the expression level of the recombinant haFGF was about 80mg/L. The expressed haFGF was purified to identity by heparin affinity chromatography and the recovery rate of haFGF was 65%. The specific activity of the purified haFGF was ED 50 4.6ng/ml.The characters of recombinant haFGF was identical to that of natural one. |
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| Keywords: | haFGF Cloning and expression E.coli. Purification Identification |
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