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| 拟南芥Antiquitin基因的原核表达和生物信息学分析 | |
| 引用本文： | 安瑞,魏少华,范三红,单丽伟.拟南芥Antiquitin基因的原核表达和生物信息学分析[J].西北植物学报,2009,29(12). |
| 作者姓名： | 安瑞 魏少华 范三红 单丽伟 |
| 作者单位： | 1. 西北农林科技大学,生命科学学院,陕西杨陵,712100 2. 西北农林科技大学,生命科学学院,陕西杨陵,712100;陕西省农业分子生物学重点实验室,陕西杨陵,712100 3. 陕西省农业分子生物学重点实验室,陕西杨陵,712100 |
| 基金项目： | 国家自然科学基金，西北农林科技大学优秀人才基金 |
| 摘 要： | 将拟南芥Antiquitin基因重组到原核表达载体pMAL-c4x和pET41中,在T7 Express菌株中诱导表达,经Amylose和Ni-NTA亲和层析柱纯化获得重组蛋白.SDS-PAGE结果表明:MBP和His-tag融合的拟南芥Antiquitin主要以可溶性形式存在,表达量分别占细胞总蛋白的25.1%和39.4%.以乙醛和NAD~+为底物测定融合蛋白活性,结果显示:His-tag融合的Antiquitin具有醛脱氢酶活性,比活力为8.98 U/mg,乙醛的表观K_m和V_(max)值分别为0.98 mmol/L和12.75 U/mg.序列比对和结构预测结果显示:拟南芥Antiquitin包含该家族蛋白典型的催化结构域、NADH结合结构域和寡聚化结构域,活性中心残基为Gly238、Gly291、Glu391、Phe393. |
| 关 键 词： | 拟南芥 重组表达 亲和纯化 |
Prokaryotic Expression and Bioinformatics Analysis of Antiquitin Protein from Arabidopsis thaliana |
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| AN Rui,WEI Shao-hua,FAN San-hong,SHAN Li-wei.Prokaryotic Expression and Bioinformatics Analysis of Antiquitin Protein from Arabidopsis thaliana[J].Acta Botanica Boreali-Occidentalia Sinica,2009,29(12). | |
| Authors: | AN Rui WEI Shao-hua FAN San-hong SHAN Li-wei |
| Institution: | AN Rui1,WEI Shao-hua1,FAN San-hong1,2,SHAN Li-wei1,2(1 College of Life Sciences,Northwest Agriculture , Forestry University,Yangling,Shaanxi 712100,China,2 Key Laboratory for Molecular Biology of Agriculture of Shaanxi Province,China) |
| Abstract: | The encoding sequence of Antiquitin protein from Arabidopsis thaliana was cloned into pMAL-c4x and pET41 vector.Fusion proteins were expressed in E.coli strain T7 Express and purified by amylose resin column and Ni-NTA resin column.Based on the results of SDS-PAGE,the yield of fusion protein,which tagged by MBP and His6,accounts for 25.1% and 39.4% of the total bacterial protein,respectively.The aldehyde dehydrogenase activity of the purified protein was determined using acetaldehyde and NAD+ substrates.The... |
| Keywords: | Antiquitin NADH Arabidopsis thaliana Antiquitin recombinant expression afinity purification NAD~+ |
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