| 拟南芥FT基因原核表达载体的构建、表达和蛋白纯化 |
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| 引用本文： | 任霞,吴忠义,黄丛林,张秀海. 拟南芥FT基因原核表达载体的构建、表达和蛋白纯化[J]. 生物技术通报, 2010, 0(3) |
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| 作者姓名： | 任霞 吴忠义 黄丛林 张秀海 |
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| 作者单位： | 1. 首都师范大学生命科学学院,北京100048;北京市农林科学院北京农业生物技术研究中心,北京,100097
2. 北京市农林科学院北京农业生物技术研究中心,北京,100097 |
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| 基金项目： | 国家"863"项目，北京市园林绿化局菊花育种创新平台 |
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| 摘 要： | FT基因是植物成花素,在植物的开花调控中起着重要作用。构建了用于原核表达的FT-eGFP表达载体,并在大肠杆菌BL21中进行重组蛋白的诱导表达。从IPTG诱导浓度、诱导时间和诱导温度等方面进行了细致的分析,最终建立了FT-eGFP融合蛋白诱导表达的优化体系:当菌液OD600=0.6-1.0时,采用IPTG1.0mol/L,在28℃诱导表达6h。摸索和建立了利用HisTrapKit标签,经过镍柱纯化,纯化目的蛋白的技术体系,为进一步研究FT基因在植物开花调控中的应用奠定了基础。
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| 关 键 词： | FT-eGFP融合蛋白 原核表达 蛋白纯化 |
Expression and Purification of Arabidopsis FT Gene in Prokaryotic Cells |
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| Ren Xia,Wu Zhongyi,Huang Conglin,Zhang Xiuhai. Expression and Purification of Arabidopsis FT Gene in Prokaryotic Cells[J]. Biotechnology Bulletin, 2010, 0(3) |
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| Authors: | Ren Xia Wu Zhongyi Huang Conglin Zhang Xiuhai |
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| Affiliation: | Ren Xia1,2 Wu Zhongyi2 Huang Conglin2 Zhang Xiuhai2(1College of Life Science,Capital Normal University,Beijing 100048,2Beijing Research Center of Agro-Biotechnology,Beijing Academy of Agriculture , Forestry Sciences,Beijing 100097) |
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| Abstract: | FT is a key gene in regulating the flowering time of Arabidopsis.In this research,expression vector of FT-eGFP in prokaryotic cells was constructed and the fused protein expression was induced in E.coli BL21.IPTG concentration,induction time and induction temperature were determined to set up optimal condition for fused protein expression purification.When OD600 of E.coli liqulid culture was 0.6-1.0,final concentration of 1.0 mol/L IPTG was applied into the culture to induce fused protein followed by incuba... |
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| Keywords: | FT-eGFP fusion protein Prokaryotic expression Protein purification |
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