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。
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.
出处
《华北农学报》
CSCD
北大核心
2013年第1期117-122,共6页
Acta Agriculturae Boreali-Sinica
基金
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)