食品科学 ›› 2017, Vol. 38 ›› Issue (16): 10-16.doi: 10.7506/spkx1002-6630-201716002

• 生物工程 • 上一篇    下一篇

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

何小用,王大明,孙文敬,崔凤杰,钱静亚,齐向辉,余泗莲   

  1. (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)
  • 出版日期:2017-08-25 发布日期:2017-08-18
  • 基金资助:
    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)

Cloning, Expression and Bioinformatic Analysis of 2-Ketogluconate Kinase Gene from Pseudomonas plecoglossicida

HE Xiaoyong, WANG Daming, SUN Wenjing, CUI Fengjie, QIAN Jingya, QI Xianghui, YU Silian   

  1. (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)
  • Online:2017-08-25 Published:2017-08-18

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%。

 

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

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.

Key words: Pseudomonas plecoglossicida, 2-ketegluconate kinase (KguK), kguK gene, cloning, expression, bioinformatics

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