十字花科植物DFR蛋白的生物信息学分析
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摘要
采用生物信息学的方法分析已经在GenBank上注册十字花科植物的DFR基因序列及相应氨基酸序列,并对其理化性质、结构特征、系统进化关系等进行预测。结果表明,十字花科植物的DFR蛋白大部分都有6个外显子,分子质量36 481.89～43 129.21 Da,大多数蛋白亚细胞定位于叶绿体中;除了BnaDFRA402,其他的DFR蛋白均具有酶活性部位、NADP结合位点和底物特异结合位点,这些保守区域形成了5个motifs;每个DFR蛋白均有多个磷酸化位点,以Ser为主,以Thr和Tyr磷酸化为辅;二级结构均由α-螺旋、β-转角、延伸链和无规则卷曲组成,其中α-螺旋为主要结构元件。
The DFR gene sequences and corresponding amino acid sequences registered on GenBank in Cruciferae plants were analyzed by means of bioinformatics, their physical and chemical properties,structural characteristics and phylogenetic relationships were predicted. The results showed that most of DFR proteins in Cruciferae plants had six exons with molecular weight ranging from 36 481.89 Da to 43 129.21 Da.Most of the protein subcells were located in chloroplasts. Except BnaDFRA402, other DFR proteins had enzymatic activity sites, NADP binding sites and substrate specific binding sites, which formed five motifs. Each DFR protein had multiple phosphorylation sites,in which Ser was the dominant and Thr and Tyr were the minor. The secondary structures were composed of alpha helix, beta turn, extended strand and random coil, and alpha helix was the main structural element.
The DFR gene sequences and corresponding amino acid sequences registered on GenBank in Cruciferae plants were analyzed by means of bioinformatics, their physical and chemical properties,structural characteristics and phylogenetic relationships were predicted. The results showed that most of DFR proteins in Cruciferae plants had six exons with molecular weight ranging from 36 481.89 Da to 43 129.21 Da.Most of the protein subcells were located in chloroplasts. Except BnaDFRA402, other DFR proteins had enzymatic activity sites, NADP binding sites and substrate specific binding sites, which formed five motifs. Each DFR protein had multiple phosphorylation sites,in which Ser was the dominant and Thr and Tyr were the minor. The secondary structures were composed of alpha helix, beta turn, extended strand and random coil, and alpha helix was the main structural element.
引文
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[18] KUMAR S,STECHER G,TAMURA K.MEGA7:Molecular Evolutionary Genetics Analysis version 7.0 for bigger datasets[J].Molecular biology and evolution,2016,33:1870-1874.
[2] 杨宏霞,曲柏宏,刘振坤,等.延边苹果梨PyDFR基因的克隆及表达分析[J].北方园艺,2015(4):99-103.
[3] 周琳,王雁,任磊,等.牡丹二氢黄酮醇-4-还原酶基因PsDFR1的克隆及表达分析[J].植物生理学报,2011,47(9):885-892.
[4] TANAKA Y,SASAKI N,OHMIYA A.Biosynthesis of plant pigments:Anthocyanins,betalains and carotenoids[J].Plant J,2008,54(4):733-749.
[5] AHMED N U,PARK J I,JUNG H J,et al.Characterization of dihydroflavonol 4-reductase(DFR)genes and their association with cold and freezing stress in Brassica rapa[J].Gene,2014,550(1):46-55.
[6] 刘娟,冯群芳,张杰.二氢黄酮醇4-还原酶基因(DFR)与花色的修饰[J].植物生理学通讯,2005,41(6):715-719.
[7] 虎娟,安韶雅,林哲,等.马蔺DFR基因的克隆及生物信息学特征分析[J].北方园艺,2017(24):109-115.
[8] MEYER P,HEIDMANN I,FORKMANN G,et al.A new petunia flower color generated by transformation of a mutant with a maize gene[J].Nature,1987,330(6149):677-678.
[9] AIDA R,KISHIMOTO S,TANAKA Y,et al.Modification of flower color in torenia(Torenia fournieri Lind.)by genetic transformation[J].Plant Sci,2000,153(1):33-42.
[10] ROSATI C,SIMONEAU P,TREUTTER D,et al.Engineering of flower color in forsythia by expression of two independently-transformed dihydrofl-avonol 4-reductase and anthocyanidin synthase genes of flavonoid pathway[J].Molecular breeding,2003,12(3):197-208.
[11] FUKUSAKI E I,KAWASAKI K,KAJIYAMA S,et al.Flower color modulations of Torenia hybrida by downregulation of chalcone synthase genes with RNA interference[J].J Biotechnol,2004,111(3):229-240.
[12] KATSUMOTO Y,FUKUCHI-MIZUTANI M,FUKUI Y,et al.Engineering of the rose flavonoid biosynthetic pathway successfully generated blue-hued flowers accumulating delphinidin[J].Plant Cell Physiol,2007,48(11):1589-1600.
[13] GASTEIGER E,GATTIKER A,HOOGLAND C,et al.ExPASy:The proteomics server for in-depth protein knowledge and analysis[J].Nucleic Acids Res,2003,31(13):3784-3788.
[14] MARCHLER-BAUER A,BO Y,HAN L,et al.CDD/SPARCLE:Functional classification of proteins via subfamily domain architec-tures[J].Nucleic Acids Res,2017,45:200-203.
[15] BAILEY T L,BODéN M,BUSKE F A,et al.MEME SUITE:Tools for motif discovery and searching [J].Nucleic acids research,2009,37:202-208.
[16] BLOM N,GAMMELTOFT S,BRUNAK S.Sequence-and structure-based prediction of eukaryotic protein phosphorylation sites[J].Journal of molecular biology,1999,294(5):1351-1362.
[17] BLOM N,SICHERITZ-PONTEN T,GUPTA R,et al.Prediction of post-translational glycolsy-lation and phosphorylation of proteins from the amino acid sequence[J].Proteomics,2004,4(6):1633-1649.
[18] KUMAR S,STECHER G,TAMURA K.MEGA7:Molecular Evolutionary Genetics Analysis version 7.0 for bigger datasets[J].Molecular biology and evolution,2016,33:1870-1874.