| 组织型纤溶酶原激活剂A链与尿激酶原B链嵌合分子的构建与表达 |
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| 引用本文： | 赵春梅,张洪涛,胡美浩.组织型纤溶酶原激活剂A链与尿激酶原B链嵌合分子的构建与表达[J].中国生物化学与分子生物学报,1999,15(4):528-531. |
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| 作者姓名： | 赵春梅 张洪涛 胡美浩 |
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| 作者单位： | 北京大学蛋白质工程及植物基因工程国家重点实验室!北京100871 |
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| 基金项目： | 863 高技术项目资助 |
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| 摘 要： | 利用 Ｄ Ｎ Ａ 重组技术和定位删除技术，将组织型纤溶酶原激活剂（ｔ Ｐ Ａ）的 Ａ 链（ Ｓｅｒｌ Ｔｈｒ２６３）基因与尿激酶原（ｐｒｏ Ｕ Ｋ）的 Ｂ链（ Ｓｅｒ１３８ Ｌｅｕ４１１）基因相连，得到嵌合分子基因ｔｕ ｐａ，并在昆虫杆状病毒系统中进行表达，表达量可达 ５００ Ｉ Ｕ／ｍ ｌ．经单克隆抗体免疫亲和层析纯化细胞表达上清液，得到ｔｕ Ｐ Ａ 嵌合分子，其比活为 ２００ ０００ Ｉ Ｕ／ｍ ｇ 蛋白． Ｓ Ｄ Ｓ Ｐ Ａ Ｇ Ｅ 及 Ｗ ｅｓｔｅｒｎ ｂｌｏｔ 鉴定证明此表达产物分子量约为 ６０ ｋ Ｄ，与预期值相符．纤维蛋白平板测活及纤维蛋白亲和性分析初步证明，此嵌合分子的溶纤活性与ｐｒｏ Ｕ Ｋ 相近，而纤维蛋白亲和性高于 ｐｒｏ Ｕ Ｋ．
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| 关 键 词： | 纤溶酶原激活剂 嵌合分子 |
| 收稿时间： | 1999-08-20 |
Construction and Expression of a Novel Chimeric Protein Consisting the A Chain of Tissue type Plasminogen Activator and the B Chain of Pro urokinase |
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| ZHAO Chunmei,ZHANG Hongtao,HU Meihao.Construction and Expression of a Novel Chimeric Protein Consisting the A Chain of Tissue type Plasminogen Activator and the B Chain of Pro urokinase[J].Chinese Journal of Biochemistry and Molecular Biology,1999,15(4):528-531. |
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| Authors: | ZHAO Chunmei ZHANG Hongtao HU Meihao |
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| Institution: | (National Laboratory of Protein Engineering and Plant Genetic Engineering,Peking University,Beijing\ 100871 |
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| Abstract: | A hybrid cDNA tu pa,which contained Serl Thr263 of tissue type plasminogen activator(t PA)and Ser138 Leu411 of pro urokinase(pro UK)was constructed via recombinant DNA technology and site directed mutagenesis.And it was expressed in the baculovirus system.The activity on fibrin plate of the cell culture was 500 IU/ml.The cell culture was purified by immuno affinity chromatography.SDS PAGE and Western blot showed that the molecular weight of tu PA was about 60 000.The specific activity of tu PA was 200 000 IU/mg protein.The fibrin affinity specificity of tu PA was much higher than that of pro UK. |
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| Keywords: | Plasminogen activator Chimeric molecule |
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