National Institute of Health Sciences, 1-18-1, Kamiyoga, Setagaya-ku, Tokyo 158-8501, Japan.
Regul Toxicol Pharmacol. 2010 Dec;58(3 Suppl):S30-5. doi: 10.1016/j.yrtph.2010.05.010. Epub 2010 May 25.
The 2D-DIGE (2-Dimensional Fluorescence Difference Gel Electrophoresis) method was applied to proteomic phenotyping of natural variants in 10 varieties of rice (Nipponbare, Koshihikari, Sasanishiki, Akitakomachi, Hitomebore, Hinohikari, Kasalath, Rexark, Bleiyo, Cho-ko) from the world rice collection (WRC) in the GenBank of the National Institute of Agrobiological Sciences (NIAS), Japan. Salt-soluble protein extracts of Nipponbare brown rice were labeled with Cy2 fluorochrome and used as an internal standard. Protein extracts from nine other rice varieties were labeled with Cy3 or Cy5 fluorochrome and applied to 2D-PAGE (13cm gel length) analysis. Approximately 700 spots of rice proteins were observed. Fluorescence intensities of each of these spots for the nine rice varieties were expressed as relative ratios to that of Nipponbare. Statistical analysis revealed the spot numbers of five Japanese rice varieties above threshold five (relative ratio of each variety to Nipponbare exceeded 5-fold or was less than 1/5) to be less than three, while those of four varieties from other countries were more than five (especially, Kasalath which was 29 and Bleiyo which was 23). The 2D-DIGE method seems to be useful for analyzing natural varieties of different cultivars and also for comparing the expression of allergen proteins.
2D-DIGE（二维荧光差异凝胶电泳）方法被应用于从日本农业生物资源研究所（NIAS）的 GenBank 中世界水稻收藏（WRC）的 10 个水稻品种（日本晴、越光、佐竹、出羽锦、响、日之光、Kasalath、Rexark、Bleiyo、朝来）的天然变异的蛋白质组学表型分析。Nipponbare 糙米的盐溶性蛋白质提取物用 Cy2 荧光染料标记，并用作内部标准。其他 9 个水稻品种的蛋白质提取物用 Cy3 或 Cy5 荧光染料标记，并应用于 2D-PAGE（13cm 凝胶长度）分析。观察到约 700 个水稻蛋白斑点。这些斑点中每个斑点的荧光强度表示为与日本晴的相对比值。统计分析显示，五种日本水稻品种的斑点数低于阈值五（每种品种与日本晴的相对比值超过 5 倍或小于 1/5）的数量少于三个，而来自其他国家的四个品种的斑点数则超过五个（特别是，Kasalath 为 29，Bleiyo 为 23）。2D-DIGE 方法似乎可用于分析不同品种的天然变异体，也可用于比较过敏原蛋白的表达。
