| 人t-PA P区在大肠杆菌中表达及其活性研究 |
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| 引用本文： | 赵月娥,郜金荣,叶林柏,徐进平,刘源洁,吴正辉.人t-PA P区在大肠杆菌中表达及其活性研究[J].武汉大学学报(理学版),2001,47(2):209-212. |
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| 作者姓名： | 赵月娥 郜金荣 叶林柏 徐进平 刘源洁 吴正辉 |
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| 作者单位： | 武汉大学 病毒研究所, |
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| 基金项目： | 湖北省科委重点项目(00151049) |
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| 摘 要： | 用PCR方法获得编码人组织型纤溶酶原激活物（tissure-typeplasminogenactivator，t-PA）C端P区肽段的DNA，并经测序证实，长810bp，含有全部的t-PA的丝氨酸蛋白酶编码序列.将这段序列克隆到表达载体pQE30中转化大肠杆菌JM109菌株,经SDS-PAGE检测转化菌株中表达出分子量约29×103的蛋白，用纤维平板法测定激活纤溶活性，结果表明表达的t-PA蛋白C端P区具有很强的激活纤溶蛋白酶原的活性.证实t-PA蛋白C端P区的酶结构区的功能是独立的,其生物活性不受其它区域影响．
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| 关 键 词： | t-PA P区 表达 活性 |
| 文章编号： | 0253-9888(2001)02-0209-04 |
| 修稿时间： | 2000年6月25日 |
Expression and Purification of t-PA P Region and Its Activity |
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| ZHAO Yue-e,GAO Jin-rong,YE Lin-Bai,Xu Jin-ping,LIU Yuan-jie,WU Zheng-hui.Expression and Purification of t-PA P Region and Its Activity[J].JOurnal of Wuhan University:Natural Science Edition,2001,47(2):209-212. |
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| Authors: | ZHAO Yue-e GAO Jin-rong YE Lin-Bai Xu Jin-ping LIU Yuan-jie WU Zheng-hui |
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| Abstract: | The DNA fragment encoding t-PA P region protein was obtained by PCR,using plasmid pTAhtpa(F) harborong the full length t-PA cDNA as template. This P region fragmemt was cloned into pQE30 and transformed to JM109 strain.P region proteins fused to 6 His peptides at its N terminus with molecular weight of 29×103 were expressed in JM109 induced by IPTG.The expressed protein were purified by Ni-NTA-agarose and were studied its activity by fibrin plate activity assay(FPAA).The results show that the function of t-PA P region is indepent for other regions of t-PA. and it has significant activity to stimulate plasminogen. |
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| Keywords: | t-PA P region expression activity |
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