变形假单胞菌葡萄糖酸脱氢酶操纵子的克隆与分析  

Cloning and bioinformatic analysis of gluconate dehydrogenase operon from Pseudomonas plecoglossicida

作　　者：宋玉玲 孙雷 孙文敬[1,3] 王大明[2,3] 崔凤杰[1,3] 齐向辉[1] SONG Yuling;SUN Lei;SUN Wenjing;WANG Daming;CUI Fengjie;QI Xianghui(School of Food and Biological Engineering,Jiangsu University,Zhenjiang 212013;School of Biotechnology,Jiangnan University,Wuxi 214122;Jiangxi Dexing Parchn Sodium Isovitamin C Co.,Ltd.,Dexing 334221)

机构地区：[1]江苏大学食品与生物工程学院,镇江212013 [2]江南大学生物工程学院,无锡214122 [3]江西省德兴市百勤异VC钠有限公司,德兴334221

出　　处：《食品科技》2019年第7期43-49,共7页Food Science and Technology

基　　金：国家自然科学基金项目(31571885);国家高技术研究发展计划项目(2012AA022103);“十三五”国家重点研发计划重点专项(2017YFC1600806);江苏高校优势学科建设工程项目(PAPD);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);德兴市科技计划项目(德科发[2015]44号)

摘　　要：该研究旨在明确变形假单胞菌JUIM01葡萄糖酸脱氢酶操纵子的结构及其基本的生物学信息。采用LA-PCR技术,从菌株JUIM01中克隆葡萄糖酸脱氢酶操纵子(gad操纵子)的全长序列;利用在线生物信息学工具,对克隆的序列进行分析;通过对变形假单胞菌膜结合葡萄糖酸脱氢酶的组成和结构分析,初步验证克隆和序列分析的结果。研究结果表明,变形假单胞菌gad操纵子具有3个结构基因gndS、gndL和gndC,分别编码膜结合葡萄糖酸脱氢酶的3个亚基。变形假单胞菌gndS基因与解淀粉欧文氏菌CFBP1430葡萄糖酸脱氢酶小亚基的编码基因的序列一致性为59.89%,其编码的蛋白属于gluconate_2-dh3 superfamily,为跨膜蛋白;基因gndL与铜绿假单胞菌PAMH19葡萄糖酸脱氢酶大亚基的编码基因的序列一致性为77.68%,其编码的蛋白属于GMC_oxred_C superfamily,为跨膜蛋白;基因gndC与香茅醇假单胞菌P3B5葡萄糖脱氢酶中亚基的编码基因的序列一致性达85.31%,其编码的蛋白属于CccA superfamily。该结果为变形假单胞菌葡萄糖酸脱氢酶的转录调控机制研究提供了一定的理论基础。The present study aimed to elucidate the structure and bioinformatics of the gluconate dehydrogenase operon(gad operon)from Pseudomonas plecoglossicida JUIM01 by cloning its full-length sequence with LA-PCR and analyzing with bioinformatics tools.The cloned sequence and analysis results were primarily verified by analyzing the components and structure of membrane-bound gluconate dehydrogenase of Pseudomonas plecoglossicida JUIM01.The results showed that the gad operon from P.plecoglossicida consisted of three structural genes including gndS,gndL and gndC,which encoding three subunits of membrane-bound gluconate dehydrogenase,respectively.The gene gndS encoded a transmembrane protein belonging to gluconate_2-dh3 superfamily with 59.89%of sequence homology of small subunit of gluconate dehydrogenase from Erwinia amylovora CFBP1430.Similarly,the gene gndL encoded a transmembrane protein belonging to GMC_oxred_C superfamily and had a 77.68%sequence homology of the large subunit of gluconate dehydrogenase from P.aeruginosa PAMH19.While the gene gndC encoded a CccA superfamily protein with a 85.31%sequence homology of the large subunit of gluconate dehydrogenase from P.citronellolis.These results might provide a theoretical basis for investigating the transcriptional regulation mechanism of gluconate dehydrogenase from P.plecoglossicida.

关 键 词：变形假单胞菌 葡萄糖酸脱氢酶 操纵子 克隆 生物信息学 

分 类 号：TS201.3[轻工技术与工程—食品科学]