明日叶二氢黄酮醇4-还原酶(AkDFR)基因的克隆及生物信息学分析  

Cloning and Bioinformatics Analysis of DFR Gene from Angelica keiskei

作　　者：刘峰 夏丽芝[2] 吴立威[1] 阮盈盈 王志江 Liu Feng;Xia Lizhi;Wu Liwei;Ruan Yingying;Wang Zhijiang(College of Landscape and Ecology,Ningbo City College of Vocational Technology,Ningbo,315100;College of Landscape Architecture and Water Conservancy Engineering,Wenzhou Vocational College of Science and Technology,Wenzhou,325006;School of Marine Sciences,Ningbo University,Ningbo,3152111;Department of Pharmaceutical Engineering,Zhejiang Pharmaceutical College,Ningbo,315100)

机构地区：[1]宁波城市职业技术学院景观生态学院,宁波315100 [2]温州科技职业学院园林与水利工程学院,温州325006 [3]宁波大学海洋学院,宁波315211 [4]浙江医药高等专科学校制药工程学院,宁波315100

出　　处：《分子植物育种》2022年第6期1838-1844,共7页Molecular Plant Breeding

基　　金：宁波市公益类科技计划项目(202002N3014);宁波市自然科学基金项目(2019A610407)共同资助。

摘　　要：二氢黄酮醇4-还原酶(DFR)基因是类黄酮合成途径的关键酶之一,在植物花色形成和黄酮物质合成中起着重要的作用。根据明日叶转录组测序得到的序列设计引物,以明日叶叶片cDNA为模板,通过RT-PCR方法克隆得到明日叶DFR基因的完整序列,并对其进行生物信息学和组织特异性分析。结果表明明日叶的AkDFR基因开放阅读框(ORF)有1 131 bp,编码376个氨基酸,蛋白大小为42.42 kD,等电点为5.83。用DNAman同源序列比对结果表明,明日叶的AkDFR基因与胡萝卜、杜鹃的DFR基因同源性均在80%以上。生物信息学分析AkDFR基因没有信号肽和跨膜区域,亲水系数为负数,可能为亲水蛋白。α-螺旋和无规则卷曲是DFR的主要二级结构元件。AkDFR基因在明日叶的不同组织中均有表达,其中叶片中的表达量最高。明日叶AkDFR基因的克隆及其生物信息学分析将为提高明日叶黄酮类化合物提供理论依据。Dihydroflavonol 4-reductase(DFR) gene is one of the key enzymes in the flavonoid synthesis pathway,and plays an important role in color formation and flavonoid synthesis in plants.The primers were designed according to the transcriptome sequence of Angelica keiskei,using cDNA of Angelica keiskei leaves as template,and the complete sequence of DFR gene in Angelica keiskei was cloned by RT-PCR method,and the bioinformatics and tissue specificity analysis were carried out.The results showed that the open reading frame(ORF) of AkDFR gene was 1 131 bp,encoding 376 amino acids,with a protein relative molecular weight of 42.42 kD and an isoelectric point of 5.83.The DNAman homology sequence alignment showed that AkDFR gene of Angelica keiskei had more than 80%homology with DFR gene of carrot and rhododendron.Bioinformatics analysis showed that AkDFR gene had no signal peptide and transmembrane region,and the hydrophilic coefficient was negative,which might be a hydrophilic protein.α-helix and random coil are the main secondary structural elements of DFR.AkDFR gene was expressed in different tissues of Angelica keiskei,with the highest expression level in leaves.The cloning and bioinformatics analysis of AkDFR gene will provide theoretical basis for improving flavonoids in Angelica keiskei.

关 键 词：明日叶(Angelica keiskei) 二氢黄酮醇4-原酶(DFR)基因 生物信息学分析 黄酮类化合物 

分 类 号：S567.239[农业科学—中草药栽培]