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。
关键词:
小麦,
磷脂酶D&alpha,
基因克隆,
生物信息学
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．
Key words:
Wheat,
Phospholipase Dα,
Gene cloning,
BioinfoRmatics
中图分类号:
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.
WANG Jun-bin, LI Ming, DING Bo, BAO Shu-guang, WANG Rui, XIE Xiao-dong. Cloning and Sequence BioinfoRmatics Analysis of Phospholipase Dα Gene fRom Wheat[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2013, 28(1): 117-122. doi: 10.3969/j.issn.1000-7091.2013.01.022.