Beom Hye-Rang, Kim Jin Sun, Jang You-Ran, Lim Sun-Hyung, Kim Chang-Kug, Lee Choung Keun, Lee Jong-Yeol
National Institute of Agricultural Science, RDA, Jeonju, 54874 Republic of Korea.
3 Biotech. 2018 Jan;8(1):56. doi: 10.1007/s13205-017-1081-2. Epub 2018 Jan 5.
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 - locus and the corresponding haplotype was -(-). Six spots were identified in the - locus and the corresponding haplotypes were - and - (-). Eight spots were identified in the - locus and the corresponding haplotypes were -, -, -, -, and - (-), 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: -, -, -, -, -, -, and -. Taken together, except for - and -, 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.
尽管已经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与其蛋白质产物之间的全面比较。本研究旨在将作者先前研究中从韩国小麦品种“Jokyoung”中分离出的43个基因与其蛋白质产物联系起来。使用二维凝胶电泳(2-DGE)分离蛋白质,并在基因单倍型水平上通过串联质谱(MS/MS)进行鉴定。通过对17个蛋白点的MS/MS分析,在-位点鉴定出2个点,相应的单倍型为-(-)。在-位点鉴定出6个点,相应的单倍型为-和-(-)。在-位点鉴定出8个点,相应的单倍型为-、-、-、-和-(-),还有1个点被γ-醇溶蛋白污染。系统发育分析以及核苷酸和氨基酸序列比对将43个基因中的35个归为7种单倍型:-、-、-、-、-、-和-。综上所述,除了未分离出的-和-,利用蛋白质组学工具和系统发育分析在基因单倍型水平上完成了每个基因与其相应蛋白质产物的关联。