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| 肾癌相关基因克隆——肾癌cDNA消减文库的构建 | |
| 引用本文： | 张强,毛泽彬,张志文,辛殿旗,郭应禄.肾癌相关基因克隆——肾癌cDNA消减文库的构建[J].生物化学与生物物理进展,2000,27(3):301-304. |
| 作者姓名： | 张强 毛泽彬 张志文 辛殿旗 郭应禄 |
| 作者单位： | 1. 北京医科大学第一临床医院,泌尿外科研究所,北京,100034 2. 北京医科大学生物化学与分子生物学系 3. 北京医科大学生理系 |
| 基金项目： | 国家自然科学基金资助项目（39870841）. |
| 摘 要： | 应用抑制性消减杂交技术，构建人肾癌与正常肾差异表达的cDNA消减文库.分别从肾癌及正常肾细胞系中提取poly(A)+RNA，依次合成单链及双链cDNA，经酶切成平均大小为400～600 bp的片段，将肾癌cDNA分为两组，分别与两种不同的接头衔接，再与正常肾cDNA进行两次消减杂交及两次抑制性PCR后，将产物与T/A载体连接构建成功cDNA消减文库，并转染大肠杆菌进行文库扩增.构建成功具有高消减效率的人肾癌cDNA消减文库，非特异性cDNA片段被有效地消减，特异表达的cDNA得到富集.文库扩增后得到6 500个克隆，随机挑取350个制备质粒，酶切分析均得到400～600 bp插入片段.所构建的人肾癌cDNA消减文库为进一步大批量筛选、克隆肾癌特异性表达的未知新基因奠定了基础. |
| 关 键 词： | 肾肿瘤，癌，抑制性消减杂交，文库 |
| 收稿时间： | 1999/6/17 0:00:00 |
| 修稿时间： | : |
Cloning of Renal Cell Carcinoma Relation Gene:Construction of a cDNA Subtractive Library of Human Renal Cell Carcinoma and Its Significance |
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| ZHANG Qiang,MAO Ze-Bin,ZHANG Zhi-Wen,XIN Dian-Qi and GUO Ying-Lu.Cloning of Renal Cell Carcinoma Relation Gene:Construction of a cDNA Subtractive Library of Human Renal Cell Carcinoma and Its Significance[J].Progress In Biochemistry and Biophysics,2000,27(3):301-304. | |
| Authors: | ZHANG Qiang MAO Ze-Bin ZHANG Zhi-Wen XIN Dian-Qi and GUO Ying-Lu |
| Institution: | Urological Department of the First Teaching Hospital, Beijing Medical University, Beijing 100034, China;Urological Department of the First Teaching Hospital, Beijing Medical University, Beijing 100034, China;Urological Department of the First Teaching Hospital, Beijing Medical University, Beijing 100034, China;Urological Department of the First Teaching Hospital, Beijing Medical University, Beijing 100034, China;Urological Department of the First Teaching Hospital, Beijing Medical University, Beijing 100034, China |
| Abstract: | |
| Keywords: | kidney neoplasms carcinoma suppression subtractive hybridization library |
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