Higginbotham J W, Smith J S, Smith O S
Pioneer Hi-Bred International, Inc., Department of Biotechnology, Johnston, IA.
Electrophoresis. 1991 Jun;12(6):425-31. doi: 10.1002/elps.1150120608.
Two-dimensional electrophoresis and fluorography of [35S]methionine labeled maize germinated embryo proteins were performed at Cold Spring Harbor Laboratory. Fluorographs of 63 gels representing 37 inbred lines were subsequently scanned and spot-detected at Protein and DNA Imageware Systems (Huntington Station, NY). The digitized images were then matched with the aid of PDQUEST-II computer software. Over 1500 different protein spots were included in the resulting dataset. The optical density data were normalized to parts per million, then transformed to their natural logarithms. Analyses of variance were performed on each spot in order to select for further study those spots with most of their variation partitioned among inbred lines rather than within inbred lines. Using this method of spot selection, over 100 protein spots were included in the set of spots which display significant differences among inbred lines of maize.
在冷泉港实验室对用[35S]甲硫氨酸标记的玉米萌发胚蛋白进行了二维电泳和荧光自显影。随后，在蛋白质和DNA图像分析系统（纽约亨廷顿站）对代表37个自交系的63块凝胶的荧光自显影片进行了扫描和斑点检测。然后借助PDQUEST-II计算机软件对数字化图像进行匹配。最终的数据集中包含了1500多个不同的蛋白质斑点。光密度数据被归一化为百万分之一，然后转换为自然对数。对每个斑点进行方差分析，以便选择那些大部分变异存在于自交系之间而非自交系内部的斑点进行进一步研究。使用这种斑点选择方法，玉米自交系间显示出显著差异的斑点组中包含了100多个蛋白质斑点。
