| SUMO融合系统高效表达可溶性鼠源FGF-21及其活性的研究 |
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| 引用本文： | 任桂萍,姜媛媛,刘铭瑶,孙国鹏,叶贤龙,李德山.SUMO融合系统高效表达可溶性鼠源FGF-21及其活性的研究[J].东北农业大学学报,2009,40(5). |
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| 作者姓名： | 任桂萍 姜媛媛 刘铭瑶 孙国鹏 叶贤龙 李德山 |
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| 作者单位： | 1. 东北林业大学博士后工作站,哈尔滨,150040;黑龙江省农科院博士后工作站,哈尔滨,150086;东北农业大学生命科学学院,哈尔滨,150030
2. 东北农业大学生命科学学院,哈尔滨,150030 |
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| 基金项目： | 黑龙江省科技厅重点攻关项目 |
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| 摘 要： | 利用SUMO表达载体在大肠杆菌中高效可溶性表达鼠源成纤维细胞生长因子(FGF-21)成熟蛋白,并研究其调节血糖的功能。将鼠源FGF-21成熟蛋白基因亚克隆至SUMO表达载体上,在大肠杆菌Rosetta(DE3)中诱导表达,并用镍离子螯合柱(Ni-NTA)纯化重组蛋白,透析后利用SUMO蛋白酶I切割融合蛋白,获得纯度较高的成熟蛋白。将3T3-L1成纤维细胞分化成脂肪细胞,经微量化的GOD-POD法检测培养基中葡萄糖含量,统计学分析葡萄糖消耗率。结果表明,在SUMO表达体系中鼠源FGF-21成熟蛋白基因以可溶形式表达,且可被SUMO蛋白酶I有效地切割,获得纯度较高的成熟蛋白。与未经处理的脂肪细胞对照组相比,经鼠源FGF-21成熟蛋白处理后脂肪细胞对葡萄糖的摄取利用显著增加,残存在培养基中的葡萄糖含量明显减少;两样本比较的t检验,P<0.001,差异极显著。
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| 关 键 词： | SUMO SUMO蛋白酶I 分子伴侣 鼠源FGF-21 葡萄糖吸收 |
Utilization of SUMO expression vector to efficiently express mouse FGF-21 and studying of its bioactivity |
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| REN Guiping,JIANG Yuanyuan,LIU Mingyao,SUN Guopeng,YE Xianlong,LI Deshan.Utilization of SUMO expression vector to efficiently express mouse FGF-21 and studying of its bioactivity[J].Journal of Northeast Agricultural University,2009,40(5). |
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| Authors: | REN Guiping JIANG Yuanyuan LIU Mingyao SUN Guopeng YE Xianlong LI Deshan |
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| Institution: | REN Guiping1,2,3,JIANG Yuanyuan3,LIU Mingyao3,SUN Guopeng3,YE Xianlong3,LI Deshan3 (1. Northeast Forestry University Postdoctoral Station,Harbin 150040,China,2. Provincial Academy of Agricultural Sciences of Heilongjiang Postdoctoral Station,Harbin 150086,3. College of Life Sciences,Northesat Agricultural University,Harbin 150030,China) |
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| Abstract: | The objective of the paper is to utilize SUMO expression vector to efficiently express mouse fibroblast growth factor (FGF)-21. Glucose regulation activity was studied for mouse mature FGF-21 protein. The cDNA of mouse mature FGF-21 protein was subcloned into the SUMO expression vector. The recombinant plasmids were transformed into Escherichia coli Rosetta (DE3) and the expressed recombinant protein was purified by its binding column. The purified recombinant protein was dialyzed against PBS. The fusion pr... |
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| Keywords: | SUMO |
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