论文

小麦磷脂酶Dα的基因克隆及其编码序列的生物信息学分析

  • 王俊斌 ,
  • 李明 ,
  • 丁博 ,
  • 包曙光 ,
  • 王锐 ,
  • 谢晓东
展开
  • 1. 天津农学院天津-布里斯托环境变化对农作物影响研究中心,天津 300384;
    2. 天津农学院基础科学系,天津 300384
王俊斌(1979-),男,内蒙古呼和浩特人,副研究员,博士,主要从事植物逆境生理研究。

收稿日期: 2012-11-17

  网络出版日期: 2014-10-14

基金资助

国家“863”计划项目(2011AA100104);天津市应用基础与前沿技术研究计划项目(11JCYBJC09100);国家转基因新品种培育重大专项(2009ZX08009-084B)

Cloning and Sequence BioinfoRmatics Analysis of Phospholipase Dα Gene fRom Wheat

  • WANG Jun-bin ,
  • LI Ming ,
  • DING Bo ,
  • BAO Shu-guang ,
  • WANG Rui ,
  • XIE Xiao-dong
Expand
  • 1. Tianjin-BRistol ReseaRch CenteR foR the Effects of the EnviRonment Change on CRops,AgRonomy,Tianjin AgRicultuRal UniveRsity,Tianjin 300384,China;

    2. DepaRtment of Basic Sciences,Tianjin AgRicultuRal UniveRsity,Tianjin 300384,China

Received date: 2012-11-17

  Online published: 2014-10-14

摘要

为获得更多小麦抗逆基因资源,利用RT-PCR方法克隆了磷脂酶D基因(PLD),命名为TaPLDα,并对基因及其蛋白进行了生物信息学分析。结果表明,TaPLDα开放阅读框为2 394 bp,编码812个氨基酸残基,等电点5.30,分子量为92 kDa。序列分析显示,TaPLDα基因编码的氨基酸序列含有N端C2结构域及2个保守的活性中心。预测TaPLDα蛋白是一个亲水性稳定蛋白且定位于细胞质,其二级结构含28.82%的α-螺旋、20.07%的延伸链、6.03%的β-转角和45.07%的不规则卷曲。该蛋白不存在信号肽,无跨膜区。TaPLDα与水稻、玉米和拟南芥的PLDα氨基酸序列之间具有高度的保守性,进化分析显示,小麦PLDα序列与毒麦、水稻和玉米PLD序列亲缘关系密切。

本文引用格式

王俊斌 , 李明 , 丁博 , 包曙光 , 王锐 , 谢晓东 . 小麦磷脂酶Dα的基因克隆及其编码序列的生物信息学分析[J]. 华北农学报, 2013 , 28(1) : 117 -122 . DOI: 10.3969/j.issn.1000-7091.2013.01.022

Abstract

To obtain moRe wheat( TRiticum aestivum L. ) stRess-Response genes,a phospholipase D gene,named TaPLDα,was cloned by RT-PCR. And the pRotein of this gene was also pRedicted and studied thRough the bioinfoRmatics analysis method. The Results showed that the laRgest open Reading fRame of TaPLDα gene has 2 394 bp in length and encoded a polypeptide of 812 amino acid Residues. The estimated moleculaR weight and isoelectRic point of the putative pRotein weRe 92 kDa and 5. 30, Respectively. Sequence analysis showed that the amino acid sequence encoded by TaPLDα gene contained N-teRminal C2 domain and two HKD motifs. TaPLDα pRotein was a stable hydRophilic pRotein and located in the cytoplasm. The pRedicted secondaRy stRuctuRe composition foR the pRotein has about 28. 82% alpha helixes,20. 07% extended stRand,6 . 03% beta tuRn and 45. 07% Random coil. The TaPLDα had no signal peptide and tRansmembRane helices. CompaRed with Rice,maize and ARabidopsis,amino acid sequence encoded by TaPLDα was almost conseRved. The phylogenic tRee showed that TaPLDα was most similaR to PLDα pRotein fRom daRnel, Rice and maize.

参考文献

[1] Hanahan D J,Chaikoff I L. A new phosphoRus-containing lipids of the caRRot[J]. J Biol Chem,1947,168: 233 - 240.

[2] Wang X,Xu L,Zheng L. Cloning and expRession of phosphatidylcholine- hydRolyzing phospholipase D fRom Ricinus communis L[J]. J Biol Chem, 1994, 269: 20312 - 20317.

[3] Wang X. The Role of phospholipase D in signal tRansduction cascade [J]. Plant Physiol, 1999, 121: 456 - 462.

[4] Li M,Hong Y,Wang X. Phospholipase D- and phosphatidic acid-mediated signaling in plants[J]. Biochim Biophys Acta, 2009, 1791: 927 - 935.

[5] Qin C,Wang X. The ARabidopsis phospholipase D family: ChaRacteRization of a calcium independent and phosphatidylcholine- selective PLDζ1 with distinct RegulatoRy domains
[J]. Plant Physiol, 2002, 128: 1057 - 1068.6] Wang C,Wang X. A novel phospholipase D of ARabidopsis that is activated by oleic acid and associated with the plasma membRane[J]. Plant Physiol,2001,127: 1102 - 1112.

[7] Wang X. RegulatoRy functions of phospholipase D and phosphatidic acid in plant gRowth,development,and stRess Responses[J]. Plant Physiol,2005,139: 566 - 573.

[8] Hong Y,Zhang W,Wang X. Phospholipase D and phosphatidic acid signaling in plant Response to dRought and salinity[J]. Plant Cell EnviRon, 2010, 33: 627 - 635.

[9] BaRgmann B O,Laxalt A M, teR Riet B,et al. Multiple PLDs RequiRed foR high salinity and wateR deficit toleRance in plants[J]. Plant Cell Physiol, 2009, 50: 78 - 89.

[10] Hong Y,Pan X,Welti R, et al. Phospholipase Dα3 is involved in the hypeRosmotic Response in ARabidopsis[J]. Plant Cell, 2008, 20: 803 - 816.

[11] Sang Y,Cui D,Wang X. Phospholipase D and phosphatidic acid-mediated geneRation of supeRoxide in ARabidopsis
[J]. Plant Physiol, 2001, 126: 1449 - 1458.

[12] MishRa G,Zhang W,Deng F, et al. A bifuRcating pathway diRects abscisic acid effects on stomatal closuRe and opening in ARabidopsis[J]. Science,2006,312: 264 - 266.

[13] Mane S P,Vasquez-Robinet C,Sioson A A, et al. EaRly PLDα1-mediated events in Response to pRogRessive dRought stRess in ARabidopsis: A tRanscRiptome analysis
[J]. J Exp Bot, 2007, 58: 241 - 252.

[14] 万嗣宝,张斌,战吉宬,等. 桃果实磷脂酶Dα 基因 的电子克隆及序列分析[J]. 中国农业科学, 2009, 42 ( 5) : 1705 - 1712.

[15] Zhang W,Wan X,Hong Y, et al. Plant phospholipase D
[M]/ / Munnik D T . Lipid signaling in plants ( plant cell monogRaphs) . BeRlin: SpRingeR publishing company, 2010: 39 - 62.

[16] Ponting C P,PaRkeR P J. Extending the C2 domain family: C2s in PKCs δ,ε,η,θ,phospholipases,GAPs,and peRfoRin[J]. PRotein Sci, 1996( 5) : 162 - 166.

[17] Zheng L,KRishnamooRthi R,Zolkiewski M, et al. Distinct Ca2 + -binding pRopeRties of novel C2 domains of plant phospholipase Dα and β[J]. J Biol Chem,2000,275: 19700 - 19706.

[18] Munnik T,ARisz S A,de VRije T, et al. G pRotein activation stimulates phospholipase D signaling in plants[J]. The Plant Cell, 1995( 7) : 2197 - 2210.

[19] Sung T C,RopeR R L,Zhang Y,et al. Mutagenesis of phospholipase D defines a defines a supeRfamily including a tRans-golgi viRal pRotein RequiRed foR poxviRus pathogenicity[J]. The EMBO JouRnal,1997,16 ( 15) : 4519 - 4530.

[20] Pointing C P,KeRR I D. A novel family of phospholipase D homologues that include phospholipid synthases and putative endonucleases: Identification of duplicated Repeats and potential active site Residues[J]. PRotein Sci, 1996( 5) : 914 - 922.

[21] 时晓伟. CIMMYT 小麦品质育种策略[J]. 天津农业 科学, 2011, 17( 3) : 108 - 111.

[22] 李萌,张爱华,白雪飞,等. 甜瓜玉米黄质环氧化酶 基因的电子克隆及生物信息学分析[J]. 华北农学 报, 2011, 26( S) : 23 - 28.
文章导航

/