文献类型：期刊文献

中文题名：丁香假单胞菌番茄致病变种DC3000甘油激酶基因的克隆与功能研究

英文题名：Cloning and Functional Analysis of Glycerol Kinase Gene of Pseudomonas syringae pathovar tomato DC3000

作者：刘晓柱 李银凤 于志海 刘晓辉

第一作者：刘晓柱

机构：[1]贵州理工学院食品药品制造工程学院

第一机构：贵州理工学院食品药品制造工程学院

年份：2018

卷号：47

期号：6

起止页码：70-75

中文期刊名：河南农业科学

外文期刊名：Journal of Henan Agricultural Sciences

收录：CSTPCD;;北大核心:【北大核心2017】；CSCD:【CSCD_E2017_2018】；

基金：贵州省教育厅创新群体重大研究项目(黔教合KY字[2017]046)

语种：中文

中文关键词：丁香假单胞菌番茄致病变种DC3000;甘油激酶;基因克隆;功能分析

外文关键词：Pst DC3000;Glycerol kinase;Gene cloning;Functional analysis

摘要：为研究丁香假单胞菌番茄致病变种(Pseudomonas syringae pathovar tomato,Pst)DC3000甘油激酶(Glycerol kinase,GK)的功能,根据Gen Bank中登录的Pst DC3000 GK氨基酸序列设计引物,克隆了Pst DC3000 GK基因,对其生物信息学特征进行分析,然后构建了GK基因敲除突变体,探讨其对菌体生理特性的影响。序列分析表明,GK基因全长1 506 bp,可编码一条501个氨基酸的多肽。生物信息学分析结果显示,Pst DC3000 GK蛋白分子质量55.8 ku,等电点5.54,富含无规卷曲结构。敲除Pst DC3000 GK后,可降低菌体在基本培养基与丰富培养基中的生长速率,增加细胞中甘油含量,同时也使菌体在拟南芥叶片上的增殖能力降低。因此,Pst DC3000 GK基因参与调控菌体的生长过程,影响菌体甘油代谢。
To probe the function of glycerol kinase(GK)in Pseudomonas syringae pathovar tomato(Pst)DC3000,GK gene was cloned with the primers that were designed according to reported Pst DC3000 GK amino acid sequence in GenBank.Then,the molecular weight,isoelectric point,primary structure and secondary structure of GK protein encoded by GK gene were analyzed using bioinformatics methods.The GK gene deletion mutant was also constructed to further explore the physiological function of GK gene.Sequence analysis showed that the length of GK gene was 1 506 bp encoding 501 amino acids.Information analysis indicated that GK molecular weight was 55.8 ku,isoelectric point was 5.54,and enriched random coil.Moreover,results showed that the bacterial growth rate reduced in media,the content of intracellular glycerol increased,and the multiplication capacity decreased in leaves of Arabidopsis after deleting GK gene.Therefore,we deduced that Pst DC3000 GK gene was involved in regulating the bacterial growth progress and the glycerol metabolism.