Fluorescent labeling of plant chromosomes in suspension by FISH
Fluorescent labeling of plant chromosomes in suspension by FISH
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DOI：
10.1266/ggs.80.35
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发表时间：
2005-02-01
影响因子：
1.1
通讯作者：
Fukui, K
中科院分区：
文献类型：
--
作者：
Ma, YZ;Lee, JH;Fukui, K
通过优化DNA合成抑制剂羟基脲（HU）和微管抑制剂氟乐灵（trifluralin）处理的浓度和时间，建立了一种高效（60%以上）的黑麦和大麦分生组织细胞周期同步化方法。从黑麦和大麦根尖分生组织中匀浆制备了含有高度纯化和形态完整的染色体的染色体悬浮液。以地高辛标记的5S rDNA为探针，对黑麦染色体进行荧光原位杂交，以生物素标记的17S rDNA和着丝粒DNA为探针，对黑麦和大麦染色体进行荧光原位杂交。在染色体上感兴趣的特定区域检测到明亮的信号。结果表明，在这项研究中开发的方法是有用的选择和分选的染色体是无法区分的其他手段，使用特定的荧光标记的悬浮液中的染色体。
By optimizing the concentration and time of treatment with hydroxyurea (HU), a DNA synthesis inhibitor, and trifluralin, a microtubule inhibitor, a highly effective (over 60%) cell cycle synchronization method for rye and barley meristem cells was developed. Chromosome suspensions containing highly purified and morphologically intact rye and barley chromosomes were prepared from the meristems of their root tips by homogenization. Digoxigenin-labeled 5S rDNA was used as a probe in FISH for the rye chromosomes in the suspension, and biotin-labeled 17S rDNA and centromeric DNA were used in FISH for the rye and barley chromosome suspensions, respectively. Bright signals were detected at the specific regions of interest on the chromosomes. The results indicate that the method developed in this study is useful for selection and sorting of chromosomes that are not distinguishable by other means, using specific fluorescent labeling by FISH of the chromosomes in suspension.