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Comprehensive identification of LMW-GS genes and their protein products in a common wheat variety

机译:全面鉴定普通小麦LMW-GS基因及其蛋白产物

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摘要

Although it is well known that low-molecular-weight glutenin subunits (LMW-GS) from wheat affect bread and noodle processing quality, the function of specific LMW-GS proteins remains unclear. It is important to find the genes that correspond to individual LMW-GS proteins in order to understand the functions of specific proteins. The objective of this study was to link LMW-GS genes and haplotypes characterized using well known Glu-A3, Glu-B3, and Glu-D3 gene-specific primers to their protein products in a single wheat variety. A total of 36 LMW-GS genes and pseudogenes were amplified from the Korean cultivar Keumkang. These include 11 Glu-3 gene haplotypes, two from the Glu-A3 locus, two from the Glu-B3 locus, and seven from the Glu-D3 locus. To establish relationships between gene haplotypes and their protein products, a glutenin protein fraction was separated by two-dimensional gel electrophoresis (2-DGE) and 17 protein spots were analyzed by N-terminal amino acid sequencing and tandem mass spectrometry (MS/MS). LMW-GS proteins were identified that corresponded to all Glu-3 gene haplotypes except the pseudogenes. This is the first report of the comprehensive characterization of LMW-GS genes and their corresponding proteins in a single wheat cultivar. Our approach will be useful to understand the contributions of individual LMW-GS to the end-use quality of flour.
机译:尽管众所周知,小麦的低分子量谷蛋白亚基(LMW-GS)会影响面包和面条的加工质量,但具体的LMW-GS蛋白的功能仍不清楚。重要的是找到与单个LMW-GS蛋白相对应的基因,以了解特定蛋白的功能。这项研究的目的是将使用众所周知的Glu-A3,Glu-B3和Glu-D3基因特异性引物表征的LMW-GS基因和单倍型与单一小麦品种的蛋白质产物关联。从韩国品种Keumkang总共扩增了36个LMW-GS基因和假基因。这些包括11个Glu-3基因单倍型,两个来自Glu-A3基因座,两个来自Glu-B3基因座,和七个来自Glu-D3基因座。为了建立基因单倍型与其蛋白产物之间的关系,通过二维凝胶电泳(2-DGE)分离出谷蛋白蛋白部分,并通过N端氨基酸测序和串联质谱(MS / MS)分析了17个蛋白斑点。鉴定出除假基因外与所有Glu-3基因单倍型相对应的LMW-GS蛋白。这是对单个小麦品种中LMW-GS基因及其相应蛋白质的全面表征的首次报道。我们的方法将有助于理解单个LMW-GS对面粉最终用途质量的贡献。

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