变形假单胞菌2-酮基葡萄糖酸操纵子的克隆与分析  

Cloning and Bioinformatics Analysis of 2-Ketogluconate Utilization Operon from Pseudomonas plecoglossicida

作　　者：栾方[1] 王大明[2,3] 孙文敬[1,2] 崔凤杰[1,2] 齐向辉[1] 郭钦[1] 张晓飞[1] 李运哲[1] Luan Fang;Wang Daming;Sun Wenjing;Cui Fengjie;Qi Xianghui;Guo Qin;Zhang Xiaofei;Li Yunzhe(School of Food and Biological Engineering, Jiangsu University, Zhenjiang 212013, Jiangsu;Parchn Sodium lsovitamin C Co. Ltd, Dexing 334221, Jiangxi;School of Biotechnology, Jiangnan University, Wuxi 214122, Jiangsu)

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：《中国食品学报》2018年第4期292-298,共7页Journal of Chinese Institute Of 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);江西省科技计划项目(赣知发[2015]64号);德兴市科技计划项目(德科发[2015]44号)

摘　　要：从变形假单胞菌JUIM01中克隆2-酮基葡萄糖酸操纵子(kgu操纵子),明确其基因组成及生物学信息。根据报道的假单胞菌的基因组信息设计简并引物,采用LA-PCR技术克隆变形假单胞菌的kgu操纵子,对克隆的基因片段进行测序,在此基础上利用生物信息学方法进行分析。结果表明:克隆到的基因片段全长为6 130bp,其中包括基因ptx S、kgu E、kgu K、kgu T和kgu D,分别编码2-酮基葡萄糖酸代谢调控蛋白Ptx S、2-酮基葡萄糖酸差向异构酶、2-酮基葡萄糖酸激酶、2-酮基葡萄糖酸转运蛋白和2-酮基葡萄糖酸-6-磷酸还原酶;这些基因与铜绿假单胞菌PAO1的kgu操纵子中相应基因的序列一致性达56.15%~76.74%,然而变形假单胞菌的ptx S很可能并不位于kgu操纵子中。研究首次从变形假单胞菌中克隆到kgu操纵子,并对其进行生物信息学分析,为变形假单胞菌2-酮基葡萄糖酸代谢机理的研究奠定了基础。To clone 2-ketogluconate utilization operon（kgu operon） from Pseudomonas plecoglossicida JUIM01, and to find its genetic organization and biological information. The primers were designed on the basis of available sequences from Ps. aeruginosa, and kgu operon was amplified by LA-PCR. The cloned fragment was sequenced and analyzed by bioinformatics. The results showed that entire nucleotide sequences（6 130 bp） were involved in the catabolism of 2 KGA and revealed five open reading frames corresponding to the ptxS gene encoding 2-ketogluconate metabolism regulation protein, the kgu E gene encoding 2-ketogluconate epimerase, the kgu K gene encoding 2-ketogluconate kinase, the kgu T gene encoding 2-ketogluconate transport protein and the kgu D gene encoding 2-ketogluconate-6-phosphate reductase.These genes were predicted to share 56.15%-76.74% sequence identity with the corresponding genes from Ps. aeruginosa PAO1. Besides, the ptxS gene might not reside in the kgu operon. The kgu operon was amplified successfully from the Ps. plecoglossicida for the first time and was analyzed by the bioinformatics. The results of this study may provide a foundation for further study about the 2-ketogluconate metabolism in Ps. plecoglossicida.

关 键 词：变形假单胞菌 2-酮基葡萄糖酸操纵子 克隆 生物信息学 

分 类 号：Q78[生物学—分子生物学] Q93