| 变形假单胞菌2-酮基葡萄糖酸激酶基因的克隆、表达及生物信息学分析 |
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| 引用本文： | 何小用,王大明,孙文敬,崔凤杰,钱静亚,齐向辉,余泗莲.变形假单胞菌2-酮基葡萄糖酸激酶基因的克隆、表达及生物信息学分析[J].食品科学,2017,38(16):10-16. |
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| 作者姓名： | 何小用 王大明 孙文敬 崔凤杰 钱静亚 齐向辉 余泗莲 |
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| 作者单位： | （1.江苏大学食品与生物工程学院，江苏?镇江 212013；2.百勤异VC钠有限公司，江西?德兴 334221；3.江南大学生物工程学院，江苏?无锡 214122） |
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| 基金项目： | 国家自然科学基金面上项目（31571885）；国家高技术研究发展计划（863计划）项目（2012AA022103）；江西省科技计划项目（赣知发[2015]64号）；江西省创新团队建设计划项目（20142BCB24024）；江西省科技平台建设计划项目（2010DTZ01900）；德兴市科技计划项目（德科发[2015]44号） |
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| 摘 要： | 采用聚合酶链式反应技术,从2-酮基葡萄糖酸工业生产菌株变形假单胞菌JUIM01中克隆2-酮基葡萄糖酸激酶的全长基因kguK,并在此基础上构建了重组菌株大肠杆菌BL21(DE3)/pET-28a(+)-kguK。在异丙基-β-D-硫代半乳糖苷的诱导下,该重组菌株表达了一个特异的分子质量约为36.0 ku融合蛋白,且该蛋白的Western-Blot鉴定结果显示为阳性。生物信息学分析结果表明,该蛋白是一种由305个氨基酸残基组成的亲水性蛋白,定位于细胞质中,存在着与pfkB家族蛋白类似的保守结构域,其二级结构中α-螺旋、延伸链和无规则卷曲所占的比例分别为35.73%、12.79%和51.48%。
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| 关 键 词： | 变形假单胞菌 2-酮基葡萄糖酸激酶 kguK基因 克隆 表达 生物信息学 |
Cloning,Expression and Bioinformatic Analysis of 2-Ketogluconate Kinase Gene from Pseudomonas plecoglossicida |
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| HE Xiaoyong,WANG Daming,SUN Wenjing,CUI Fengjie,QIAN Jingya,QI Xianghui,YU Silian.Cloning,Expression and Bioinformatic Analysis of 2-Ketogluconate Kinase Gene from Pseudomonas plecoglossicida[J].Food Science,2017,38(16):10-16. |
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| Authors: | HE Xiaoyong WANG Daming SUN Wenjing CUI Fengjie QIAN Jingya QI Xianghui YU Silian |
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| Affiliation: | (1. School of Food and Biological Engineering, Jiangsu University, Zhenjiang 212013, China; 2. Parchn Sodium Isovitamin C Co. Ltd., Dexing 334221, China; 3. School of Biotechnology, Jiangnan University, Wuxi 214122, China) |
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| Abstract: | The full-length sequence of 2-ketegluconate kinase gene (kguK) from was cloned from an industrial 2KGA producer of Pseudomonas plecoglossicida JUIM01 and expressed in the recombinant strain Escherichia coli BL21(DE3)/pET-28a(+)-kguK with isopropyl β-D-1-thiogalactopyranoside induction. The specific fusion protein KguK had a molecular weight of about 36.0 ku, which was confirmed by Western-Blot. The bioinformatics analysis showed that KguK in P. plecoglossicida was a hydrophilic protein with 305 amino acid residues. It was located in the cytoplasm, having a conserved domain similar to that of the pfkB family. The predicted secondary structure contained 35.73% of α-helixes, 12.79% of extended strand and 51.48% of random coil. |
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| Keywords: | Pseudomonas plecoglossicida 2-ketegluconate kinase (KguK) kguK gene cloning expression bioinformatics |
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