Mueller G M, Wolber P K, Warren G J
DNA Plant Technology Corporation, Oakland, California 94608.
Cryobiology. 1990 Aug;27(4):416-22. doi: 10.1016/0011-2240(90)90018-y.
Antibodies raised against a synthetic peptide specifically detect ice nucleation proteins from Pseudomonas species in Western blots. In immunofluorescent staining of whole bacteria, the antibodies reveal the protein in clusters, as indicated by patches of intense fluorescence in Escherichia coli cells heterologously expressing Pseudomonas ice nucleation genes. The abundance, size, and brightness of the clusters vary considerably from cell to cell. Their varying sizes may explain the variability in activity of bacterial ice nuclei. Growth at lower temperatures produces more ice nuclei, and gives brighter and more frequent patches, than growth at 37 degrees C. The observed clustering may thus reflect formation of functional ice nucleation sites in vivo. The presence of ice nucleation protein in clusters is also correlated with alterations in cell morphology.
针对合成肽产生的抗体在蛋白质印迹法中能特异性检测来自假单胞菌属的冰核蛋白。在对完整细菌的免疫荧光染色中，抗体显示该蛋白呈簇状分布，如在异源表达假单胞菌冰核基因的大肠杆菌细胞中出现的强烈荧光斑块所示。这些簇的丰度、大小和亮度在细胞间差异很大。它们大小的变化可能解释了细菌冰核活性的变异性。与在37℃下生长相比，在较低温度下生长会产生更多的冰核，并且会出现更亮、更频繁的斑块。因此，观察到的簇状分布可能反映了体内功能性冰核位点的形成。簇中冰核蛋白的存在也与细胞形态的改变相关。
