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Proteomic analysis of low-molecular-weight glutenin subunits and relationship with their genes in a common wheat variety

机译:普通小麦品种中低分子量谷蛋白亚基的蛋白质组学分析及其与基因的关系

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摘要

Although many studies on low-molecular-weight glutenin subunit (LMW-GS) function have been reported, a comprehensive comparison between specific genes and their protein product is still lacking. This study aimed to link the 43 genes isolated from the Korean wheat variety “Jokyoung” in the authors’ previous study to their protein products. Proteins were separated using two-dimensional gel electrophoresis (2-DGE) and identified by tandem mass spectrometry (MS/MS) at the gene haplotype level. Using MS/MS analysis of 17 protein spots, two spots were identified in the Glu-A3 locus and the corresponding haplotype was GluA3-13(Glu-A3c). Six spots were identified in the Glu-B3 locus and the corresponding haplotypes were GluB3-33 and GluB3-43 (Glu-B3h). Eight spots were identified in the Glu-D3 locus and the corresponding haplotypes were GluD3-11, GluD3-21, GluD3-31, GluD3-5, and GluD3-6 (Glu-D3a), and one spot was contaminated with gamma gliadin. Phylogenetic analysis and alignment of nucleotide and amino acid sequences assigned 35 of the 43 genes to seven haplotypes: GluA3-13, GluB3-43, GluD3-11, GluD3-21, GluD3-31, GluD3-42, and GluD3-5. Taken together, except for GluB3-33 and GluD3-6, which were not isolated, linking of each gene to the corresponding protein products at the gene haplotype level was accomplished using proteomic tools and phylogenetic analysis.Electronic supplementary materialThe online version of this article (10.1007/s13205-017-1081-2) contains supplementary material, which is available to authorized users.
机译:尽管已报道了许多关于低分子量谷蛋白亚基(LMW-GS)功能的研究,但仍缺乏对特定基因及其蛋白质产物的全面比较。这项研究的目的是将作者先前研究中从韩国小麦品种“ Jokyoung”中分离的43个基因与他们的蛋白质产物联系起来。使用二维凝胶电泳(2-DGE)分离蛋白质,并通过串联质谱(MS / MS)在基因单倍型水平上进行鉴定。通过MS / MS分析17个蛋白斑点,在Glu-A3基因座中鉴定出两个斑点,相应的单倍型为GluA3-13(Glu-A3c)。在Glu-B3基因座中鉴定出六个斑点,相应的单倍型为GluB3-33和GluB3-43(Glu-B3h)。在Glu-D3基因座中鉴定出8个斑点,相应的单倍型为 GluD3 - 11 GluD3 - 21 GluD3 - 31 GluD3 - 5 GluD3 - 6 Glu - D3a ),其中一个点被伽马麦醇溶蛋白污染。系统进化分析和核苷酸和氨基酸序列的比对,将43个基因中的35个分配给七个单倍型: GluA3 - 13 GluB3 - 43 GluD3 - 11 GluD3 - 21 GluD3 - 31 GluD3 - 42 GluD3 - 5 。在一起,除了未分离的 GluB3 - 33 GluD3 - 6 ,每个基因都连接在一起使用蛋白质组学工具和系统发育分析完成了在基因单倍型水平上对相应蛋白质产品的纯化。电子补充材料本文的在线版本(10.1007 / s13205-017-1081-2)包含补充材料,可供授权用户使用。

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