| CUEDC2中CUE结构域的表达纯化及结构初探 |
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| 引用本文： | 崔佳梅,张明,张学敏,颜贤忠.CUEDC2中CUE结构域的表达纯化及结构初探[J].生物技术通讯,2011,22(4):509-512. |
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| 作者姓名： | 崔佳梅 张明 张学敏 颜贤忠 |
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| 作者单位： | 军事医学科学院,生物医学分析中心,北京,100850 |
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| 摘 要： | 目的:建立CUEDC2中CUE结构域的原核表达系统,获得13C、15N同位素标记的CUE结构域蛋白质,以用于结构生物学研究。方法:利用分子生物学方法将CUE结构域编码序列构建到pET-28a原核表达系统,表达和纯化13C、15N标记的重组蛋白;用SDS-PAGE等方法对其进行鉴定。结果:目的蛋白经SDS-PAGE和MALDI-TOF/MS检测,相对分子质量正确,圆二色谱和核磁共振波谱结果显示目的蛋白折叠良好。结论:获得了高浓度、高纯度、折叠良好的CUE结构域标记蛋白质,利于进一步的结构生物学研究。
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| 关 键 词： | CUE结构域 CUEDC2 MALDI-TOF/MS 核磁共振 结构生物学 |
Expression and Purification of CUE Domain in CUEDC2 and Preliminary Investigation on its Structure |
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| CUI Jia-Mei,ZHANG Ming,ZHANG Xue-Min,YAN Xian-Zhong.Expression and Purification of CUE Domain in CUEDC2 and Preliminary Investigation on its Structure[J].Letters in Biotechnology,2011,22(4):509-512. |
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| Authors: | CUI Jia-Mei ZHANG Ming ZHANG Xue-Min YAN Xian-Zhong |
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| Institution: | National Center of Biomedical Analysis,Beijing 100850,China |
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| Abstract: | Objective： To obtain stable-isotope 13C/15N-labeled CUE domain of CUEDC2 by constructing prokaryotic expression vector carrying DNA fragment encoding CUE domain in CUEDC2 for structural and functional studies.Methods： The coding sequence of the CUE was cloned into pET-28a prokaryotic expression system.The His-tagged CUE was expressed and purified with affinity purification technology,identified with SDS-PAGE,MALDI-TOF/MS,circular dichroism and NMR.Results： The target peptide was correctly expressed as indicated by the results from SDS-PAGE and MALDI-TOF/MS;2D 1H-15N HSQC NMR spectroscopy and circular dichroism data showed that CUE domain was well folded.Conclusion： Well folded CUE domain sample with isotope-labeling was correctly expressed with high quality and was suitable for further solution structure study. |
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| Keywords: | CUEDC2 MALDI-TOF/MS |
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