| 人生存素survivin的克隆与分泌表达 |
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| 引用本文： | 代晓华,姜琳琳,程昕,纪庆,朱惠芳,熊冬生,周圆. 人生存素survivin的克隆与分泌表达[J]. 生物技术通报, 2012, 0(5): 89-92 |
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| 作者姓名： | 代晓华 姜琳琳 程昕 纪庆 朱惠芳 熊冬生 周圆 |
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| 作者单位： | 1. 中国医学科学院北京协和医学院血液病医院(血液学研究所)实验血液学国家重点实验室,天津300020;天津市口腔医院暨南开大学附属口腔医院实验研究中心,天津300041
2. 中国医学科学院北京协和医学院血液病医院(血液学研究所)实验血液学国家重点实验室,天津,300020 |
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| 基金项目： | 高等学校博士学科点专项科研基金，国家自然科学基金，天津市自然科学基金 |
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| 摘 要： | 旨在克隆人生存素( survivin)基因,并在原核系统中进行可溶性表达.采用RT-PCR方法从人乳腺癌细胞MCF-7中克隆survivin基因,将其重组到pAYZ表达栽体中,构建带有六聚组氨酸纯化标记的人survivin基因原核表达栽体pAYZ-survivin,将该载体转化大肠杆菌16C9进行表达.结果表明,成功克隆了survivin基因并构建了重组表达载体.融合蛋白主要以可溶性状态分泌到周质腔内存在.分离提取蛋白,HiTrap金属螯合柱进行亲和层析纯化,并经Western blot验证了高纯度survivin融合蛋白的表达.
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| 关 键 词： | 生物医学工程 生存素 分泌表达 大肠杆菌 凋亡 |
Cloning and Secretory Expression of Recombinant Human Survivin Gene |
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| Dai Xiaohua , Jiang Linlin , Cheng Xin , Ji Qing , Zhu Huifang , Xiong Dongsheng , Zhou Yuan. Cloning and Secretory Expression of Recombinant Human Survivin Gene[J]. Biotechnology Bulletin, 2012, 0(5): 89-92 |
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| Authors: | Dai Xiaohua Jiang Linlin Cheng Xin Ji Qing Zhu Huifang Xiong Dongsheng Zhou Yuan |
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| Affiliation: | 1(1State Key Laboratory of Experimental Hematology,Institute of Hematology and Blood Diseases Hospital,Chinese Academy of Medical Sciences and Peking Union Medical College,Tianjin 300020;2Experimental Research Center,Tianjin Stomatological Hospital,Affiliated Dental College of Nankai University,Tianjin 300041) |
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| Abstract: | It was to clone human survivin gene and express it in a dissolute way in the prokaryotic system.The full-length human survivin gene was amplified from breast cancer cell line MCF-7 by RT-PCR,and then was cloned into pAYZ expression vector to construct the recombinat pAYZ-survivin with 6×His-tag.Escherichia coli 16C9 were transformed with the recombinant vector.Results showed the recombinant survivin protein was secreted into the periplasm of E.coli in a soluble form.After isolation and purification by HiTrap Chelating column,western blot analysis was performed to confirm the soluble expression of the recombinant protein. |
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| Keywords: | Biomedical engineerin Survivin Secretory expression Escherichia coli Apoptosis |
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