| 从包涵体中纯化重组小鼠Nanog转录因子 |
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| 引用本文： | 李军,李勇,吕长荣,窦忠英. 从包涵体中纯化重组小鼠Nanog转录因子[J]. 西北农业学报, 2009, 18(6): 1-4 |
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| 作者姓名： | 李军 李勇 吕长荣 窦忠英 |
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| 作者单位： | 李军(宁夏职业技术学院,生物工程系,宁夏银川,750002;西北农林科技大学,国家干细胞工程技术研究中心陕西分中心,陕西杨凌,712100);李勇(西北农林科技大学,国家干细胞工程技术研究中心陕西分中心,陕西杨凌,712100;宁夏大学,西部特色生物资源利用与保护教育部重点实验室,宁夏银川,750021);吕长荣,窦忠英(西北农林科技大学,国家干细胞工程技术研究中心陕西分中心,陕西杨凌,712100) |
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| 基金项目： | 国家"863"项目，教育部重大项目 |
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| 摘 要： | 利用BL21/pET-32a-Nanog表达菌株诱导表达Nanog融合蛋白,经破碎、高速离心、洗涤,获得Nanog蛋白粗制品,用8 moL/L尿素变性溶解Nanog融合蛋白,离心取上清,进行Histrap HP亲和层析纯化,复性,经SDS-PAGE和Western Blot鉴定得到了纯度为97%以上的重组小鼠Nanog蛋白.建立了一种可行的从包涵体中纯化重组小鼠Nanog蛋白的方法,为Nanog蛋白的活性检测及其基因表达调控机理的功能研究奠定基础.
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| 关 键 词： | 小鼠 包涵体 纯化 |
| 收稿时间： | 2009-06-02 |
| 修稿时间： | 2009-09-24 |
Purified Recombinant Mouse Nanog Transcription Factor from Inclusion Body |
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| LI Jun,LI Yong,L,#; Changrong and DOU Zhongying. Purified Recombinant Mouse Nanog Transcription Factor from Inclusion Body[J]. Acta Agriculturae Boreali-occidentalis Sinica, 2009, 18(6): 1-4 |
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| Authors: | LI Jun,LI Yong,L&# Changrong DOU Zhongying |
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| Affiliation: | Bioengineering Departement of Ningxia Vocational&Technical College, Yinchuan Ningxia 750002, China;Shaanxi Branch of National Stem Cell Engineering Center, Northwest A&F University, Yangling Shaanxi 712100, China;Shaanxi Branch of National Stem Cell Engineering Center, Northwest A&F University, Yangling Shaanxi 712100, China;Key Laboratory of Ministry of Education for Protection and Utilizatation of Special Biological Resources in Western China, Ningxia University, Yinchuan Ningxia 750021, China;Shaanxi Branch of National Stem Cell Engineering Center, Northwest A&F University, Yangling Shaanxi 712100, China;Shaanxi Branch of National Stem Cell Engineering Center, Northwest A&F University, Yangling Shaanxi 712100, China |
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| Abstract: | In this experiment, Nanog fusion protein was expressed by BL21/pET-32a-Nanog, and the crude Nanog protein was obtained from inclusion body by crushing, high speed centrifuging and washing. Then the inclusion body was dissolved by 8M urea and centrifuging. The supernatant, which be used to Histrap HP column gradual dialysis, renatured and concentrated, was got by centrifuging. the purity of Nanog fusion protein was analyzed by SDS-PAGE and Western blot, and it is up to 97%. The way of preparing mouse Nanog fusion protein from the inclusion body was established, which was prepared for activation detection and mechanism of gene expression and regulation. |
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| Keywords: | Nanog |
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