| 转录因子CTCF的克隆、表达及其抗体制备 |
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| 引用本文： | 何莉,朱旭东,毛志勇,朱恒奇,黄培堂. 转录因子CTCF的克隆、表达及其抗体制备[J]. 生物技术通讯, 2004, 15(5): 454-456 |
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| 作者姓名： | 何莉 朱旭东 毛志勇 朱恒奇 黄培堂 |
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| 作者单位： | 1. 军事医学科学院,生物工程研究所,北京,100071;中国药品生物制品检定所,北京,100050
2. 军事医学科学院,生物工程研究所,北京,100071 |
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| 摘 要： | 用RT-PCR方法从HeLa细胞总RNA中扩增转录因子CTCF的cDNA序列，克隆至pGEM-TEasy载体上。将CTCF蛋白N段的编码序列亚克隆至pET22b( )载体中，在大肠杆菌BL21(DE3)中诱导表达，并用镍柱进行亲和纯化。用纯化后的CTCFN段蛋白免疫家兔，获得CTCF的多克隆抗体，进而用于细胞内CTCF蛋白的免疫印迹检测。
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| 关 键 词： | 转录因子 蛋白 抗体制备 诱导表达 亚克隆 细胞内 ET 纯化 cDNA序列 总RNA |
| 文章编号： | 1009-0002(2004)05-0454-03 |
| 修稿时间： | 2004-04-29 |
Cloning, expression and antibody preparation of transcription factor CTCF |
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| HE Li,,ZHU Xu-dong,MAO Zhi-yong,ZHU Heng-qi,HUANG Pei-tang. Cloning, expression and antibody preparation of transcription factor CTCF[J]. Letters in Biotechnology, 2004, 15(5): 454-456 |
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| Authors: | HE Li ZHU Xu-dong MAO Zhi-yong ZHU Heng-qi HUANG Pei-tang |
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| Affiliation: | HE Li1,2,ZHU Xu-dong1,MAO Zhi-yong1,ZHU Heng-qi1,HUANG Pei-tang11. Institute of Biotechnology,Academy of Military Medical Sciences,Beijing 100071,2. National Institute for the Control of Pharmaceutical and Biological Products,Beijing 100050,China |
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| Abstract: | CTCF cDNA was amplified from total RNA of HeLa cells and was ligated into pGEM-T Easy plasmid. N domain of CTCF, termed CTCF-N, was expressed in E.coli BL21(DE3) using pET expression system, and then CTCF-N was purified by Ni-NTA agarose column. CTCF-N polyclonal antibody was elicited by immunizing rabbit, then purified and used to detect CTCF level by western blotting. |
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