福建农业学报2016，Vol.31Issue(7)：683-689,7.DOI:10.19303/j.issn.1008-0384.2016.07.003
刺葡萄 DFR 基因克隆及生物信息学分析
Cloning and Bioinformatics of DFR Gene in Vitis davidii Foёx
摘要
Abstract
The specific primers of open reading frame (ORF)of dihydroflavonol (DFR)gene in brier grapes (Vitis davidii Foёx.)were designed according to the CDS template of the gene.DFR gene sequence was cloned using RT-PCR,andsubsequently,the genetic characteristics analyzed by bioinformatics.The 1 014 bp full-length cDNA of DRF's ORF was thus obtained.It encoded 337 amino acids,and was named V .davidii DFR 4-reductase gene (VdDFR )with GenBank accession number of KF91 5803. The predicted molecular weight of VdDFR protein was 37 593.2 Da,theoretical pI is 5.81. As a transmembrane and a hydrophilic protein,it did not belong to secretory category,had no signal peptide,and was located largely in the cytoplasm (70%).The secondary structure ofthe mixed protein was mostly random coil (52.82%).The amino acids sequence of the gene possibly contained 7 glycosylation sites,1 6 phosphorylation sites,one NAD (P)binding site,and one NAD-dependent epimerase/dehydratase (N-terminal) domain,and the gene likely belonged to the NADB _ Rossmann superfamily.The nucleotide sequences of DFR from V . davidii,V . bellula,V . amurensis , and V . vinifera were 99%homogenous;those of V .davidii and V .rotundifolia, 98% homogenous; and, those of V .davidii and Ampelopsis grossedentata , 94% homogenous.These results indicated that the DFR gene coding region was evolutionally conservative.And,the phylogenetic tree constructed based on the sequence salso reflected the same evolutionary trait of these plants.关键词
刺葡萄/二氢黄酮醇 4-还原酶/DFR 基因/克隆/生物信息学Key words
brier grape (Vitisdavidii Foёx.)/dihydroflavonol 4-reductase/DFR gene/cloning/bioinformatics分类
农业科技引用本文复制引用
赖呈纯,黄贤贵,甘煌灿,潘红,范丽华..刺葡萄 DFR 基因克隆及生物信息学分析[J].福建农业学报,2016,31(7):683-689,7.基金项目
福建省科技计划项目---省属公益类科研院所基本科研专项(2014R1015-6) （2014R1015-6）
福建省自然科学基金项目(2016J01126) （2016J01126）
