郑麦366α-醇溶蛋白基因的克隆及生物信息学分析

为研究优质强筋小麦品种郑麦366α-醇溶蛋白的组成,应用简并引物进行PCR扩增,从郑麦366中扩增得到13条核苷酸序列,其中9条序列推导的氨基酸序列具有完整的开放阅读框。进一步分析显示,克隆的9个基因(分别命名为ZM366-1—ZM366-9)编码的蛋白质均具有α-醇溶蛋白的典型结构特征,根据T-细胞毒性抗原表位数目和多聚谷氨酰胺区特征分别将ZM366-1、ZM366-2定位到6A染色体,ZM366-3、ZM366-4定位到6D染色体,ZM366-5—ZM366-9定位到6B染色体。蛋白质二级结构预测显示,克隆的9个α-醇溶蛋白都仅含有α-螺旋结构,其中B基因组α-醇溶蛋白的α-螺旋含量明显高于其他基因组。同源系统进化树分析表明,克隆的9个α-醇溶蛋白具有明显的基因组特异性。

Cloning and Bioinformatic Analysis of α-gliadin Genes in Zhengmai 366

To reveal the composition of α-gliadin protein in high quality and strong gluten wheat variety Zhengmai 366,the PCR amplification were applied to clone α-gliadin genes in Zhengmai 366 with degenerate primers.In total,13 nucleotide sequences were amplified,while the deduced amino acid sequences of nine encoded complete open reading frames.Further analysis showed that the cloned genes (named as ZM366-1-ZM366-9 respectively) had the typical characteristics of α-gliadin.According to the number of T cell stimulatory toxic epitopes and the characteristics of polyglutamine domain,ZM366-1 and ZM366-2,ZM366-3 and ZM366-4 and ZM366-5-ZM366-9 were localized to the 6A,6D and 6B chromosome respectively.Secondary structure prediction showed that all the nine α-gliadin cloned in the present study only contained α-helix structure,and the content of α-helix was significantly higher in α-gliadin of B genome than that of others.Phylogenetic tree analysis showed that the cloned α-gliadin of present study had obvious genomic specificity.