文献类型：期刊文献

中文题名：地黄己糖激酶RgHXK2基因的电子克隆及生物信息学分析

英文题名：In Silico Cloning and Bioinformatics Analysis of Hexokinase Gene RgHXK2 in Rehmannia glutinosa

作者：王晨璨[1,5] 李瑾[3] 聂双喜[1] 吕亚奎[6] 彭霄鹏[2,4,6]

第一作者：王晨璨

机构：[1]广西大学轻工与食品工程学院,广西清洁化制浆造纸与污染控制重点实验室,南宁530004;[2]齐鲁工业大学(山东省科学院),齐鲁工业大学制浆造纸科学与技术教育部重点实验室,济南250353;[3]中国林业科学研究院资源昆虫研究所,昆明650224;[4]中国林业科学研究院林业研究所,国家林业和草原局林木培育重点实验室,北京100091;[5]北京林业大学生物科学与技术学院,林木育种国家工程实验室,北京100083;[6]海南热带野生动植物园有限公司,海口571154

年份：2023

卷号：21

期号：19

起止页码：6335-6345

中文期刊名：分子植物育种

外文期刊名：Molecular Plant Breeding

收录：CSTPCD;;北大核心:【北大核心2020】；

基金：中央级公益性科研院所基本科研业务费专项(CAFYBB2022SY003);广西清洁化制浆造纸与污染控制重点实验室开放基金项目(2021KF36);制浆造纸科学与技术教育部重点实验室开放基金项目(KF201909);南京林业大学南方现代林业协同创新中心和南海系列育才计划共同资助。

语种：中文

中文关键词：地黄(Rehmannia glutinosa);RgHXK2;电子克隆;生物信息学分析

外文关键词：Rehmannia glutinosa;RgHXK2;In silico cloning;Bioinformatics analysis

分类号：R28

摘要：己糖激酶(HXK)的主要功能是在糖酵解途径中催化己糖磷酸化,是植物体内调控呼吸代谢的关键酶之一。主要催化己糖磷酸化进入糖酵解途径,可在植物体的生长发育过程中为其提供大量的能量。己糖激酶属于hexokinase-2超家族,在植物生命活动中发挥着重要的作用。为了解地黄(Rehmannia glutinosa)己糖激酶基因RgHXK2在磷酸化过程中起到的作用,该研究基于转录组测序获得的地黄己糖激酶基因ORF的部分保守序列和地黄的SRA数据库,在NCBI中进行电子克隆获得地黄RgHXK2全长cDNA序列;采用在线网上工具ProtParam、TMHMM、SOPMA、SMART、MEME、Clustal Omega以及Jalview、MegaX等软件对其进行生物信息学分析,结果显示,地黄RgHXK2基因全长1 462 bp,包含1 194 bp的开放阅读框,编码一个含有397个氨基酸、相对分子质量为43.75 kD、等电点为6.34的蛋白质。在线软件预测该蛋白含有Hexokinase-1和Hexokinase-2保守结构域,无信号肽与跨膜结构,为亲水性非分泌蛋白,主要分布于线粒体和叶绿体中,参与碳水化合物代谢、细胞葡萄糖的动态平衡以及糖酵解等三个生物反应过程。系统进化分析表明,地黄与木犀榄(Olea europaea)、一串红(Salvia splendens)、黄色猴面花(Erythranthe guttata)、紫花风铃木(Handroanthus impetiginosus)及松蒿(Phtheirospermum japonicum)、独脚金(Striga asiatica)聚为一支,其中与木犀榄HXK1蛋白的同源性最高,为90.05%。本研究为进一步揭示RgHXK2基因在地黄己糖磷酸化途径中的功能奠定基础。
Hexokinase(HXK)is one of the key enzymes related to respiration and metabolism in plants.It plays an important role in catalyzing hexose phosphorylation and making hexose enter the glycolysis pathway,and provides a large amount of energy for the growth and development of plants.Hexokinase belongs to the Hexokinese-2 super family and it is significant in plant life activities.To understand the function of Rehmannia hexokinase HXK genes in the process of its phosphorylation,we report here a complete analysis of RgHXK2 gene based on transcriptome sequencing of Rehmannia hexokinase2 gene ORF sequences of some conservative and Rehmannia SRA database,in silico cloning for radix rehmanniae HXK in NCBI gene cDNA,and adopt ProtParam,TMHMM,SOPMA,SMART,MEME,Clustal Omega,Jalview and MegaX bioinformatics analysis software to analyze it.The results showed that the cDNA full length of RgHXK2 gene in Rehmannia was 1462 bp,encodes a protein containing 397 amino acids with a relative molecular weight of 43.75 kD and a theoretical pI was 6.34.It was speculated that this protein had no transmembrane transport function and was not a secreted protein.Structural analysis based on the amino acid sequence revealed that RgHXK2 had two conserved domains,Hexokinese-1 and Hexokinese-2,no signal peptide and transmembrane structure,and was a hydrophilic non-secreted protein,mainly distributed in mitochondria and chloroplasts,which was involved in carbohydrate metabolism,cellular glucose homeostasis,and glycolysis.Phylogenetic analysis showed that Rehmannia glutinosa with Olea europaea subsp.Europaea,Salvia splendens,Erythranthe guttata,Handroanthus impetiginosus,Phtheirospermum japonicum and Striga asiatica were a cluster,and the homology of RgHXK2 and the OeHXK1 gene of Olea europaea subsp.Europaea was up to about 90.05%.This study has laid a foundation for further revealing the role of RgHXK2 gene in the hexose phosphorylation pathway of Rehmannia glutinosa.