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  

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

作　　者：何小用 王大明[2,3] 孙文敬[1,2] 崔凤杰[1,2] 钱静亚[1,2] 齐向辉[1] 余泗莲[2] HE Xiaoyong;WANG Daming;SUN Wenjing;CUI Fengjie;QIAN Jingya;QI Xianghui;YU Silian(School of Food and Biological Engineering, Jiangsu University, Zhenjiang 212013, China;Parchn Sodium Isovitamin C Co. Ltd., Dexing 334221, China;School of Biotechnology, Jiangnan University, Wuxi 214122, China)

机构地区：[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　　处：《食品科学》2017年第16期10-16,共7页Food Science

基　　金：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)

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%。The full-length sequence of2-ketegluconate kinase gene(kguK)from was cloned from an industrial2KGA producer of Pseudomonas plecoglossicida JUIM01and expressed in the recombinant strain Escherichia coli BL21(DE3)/pET-28a(+)-kguK with isopropylβ-D-1-thiogalactopyranoside induction.The specific fusion protein KguK had a molecular weight of about36.0ku,which was confirmed by Western-Blot.The bioinformatics analysis showed that KguK in P.plecoglossicida was a hydrophilic protein with305amino acid residues.It was located in the cytoplasm,having a conserved domain similar to that of the pfkB family.The predicted secondary structure contained35.73%ofα-helixes,12.79%of extended strand and51.48%of random coil.

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 

分 类 号：Q781[生物学—分子生物学]