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首页> 外文期刊>Zoological Science >Production of transgenic medaka fish carrying fluorescent nuclei and chromosomes
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Production of transgenic medaka fish carrying fluorescent nuclei and chromosomes

机译：带有荧光核和染色体的转基因高加索鱼的生产

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As with zebrafish, attention has focused on the teleost medaka Oryzias latipes as an experimental animal representative of non-mammalian vertebrates in various fields of biological science. To enable real-time analyses of the dynamics of nuclei and chromosomes in living medaka cells, we produced a transgenic medaka expressing a fusion protein between histone H2B and green fluorescent protein (GFP) under the control of a cytomegalovirus (CMV) promoter. Since the nuclei and chromosomes of transgenic medaka cells are labeled with GFP, their morphological changes can be instantly monitored throughout the mitotic cell cycle progression under a fluorescent microscope without any fixation and staining of samples. However, GFP-labeling of nuclei and chromosomes is not successful during early embryonic development until zygotic expression begins and during the meiotic cell cycle progression, because the CMV promoter does not work in these stages. In addition, histone H2B-GFP fusion proteins are expressed in an organ-specific manner; strong and ubiquitous expression occurs in cells comprising the gut and fin, whereas the expression is restricted to certain types of cells in the liver and brain. These findings suggest that the CMV-driven expression of the histone H2B-GFP transgene is modified depending on the integration site of the transgene in the genome. Nevertheless, easy and precise monitoring of cytological changes in nuclei and chromosomes in the majority of mitotic cells by using the transgenic medaka will greatly contribute to a better understanding of control mechanisms of nuclear and chromosomal behaviors in vertebrate cells.
机译：与斑马鱼一样，人们将注意力集中在硬骨，作为生物学上各个领域的非哺乳动物脊椎动物的实验动物代表。为了能够实时分析活果细胞中核和染色体的动力学，我们生产了一种转基因果，它在巨细胞病毒（CMV）启动子的控制下表达组蛋白H2B和绿色荧光蛋白（GFP）之间的融合蛋白。由于转基因花aka细胞的细胞核和染色体都用GFP标记，因此可以在荧光显微镜下在整个有丝分裂细胞周期进程中立即监测其形态变化，而无需对样品进行任何固定和染色。但是，在早期胚胎发育之前，直到合子表达开始和减数分裂细胞周期发展期间，对核和染色体的GFP标记都是不成功的，因为CMV启动子在这些阶段不起作用。另外，组蛋白H2B-GFP融合蛋白以器官特异性方式表达;在包括肠和鳍的细胞中发生强而普遍的表达，而表达仅限于肝和脑中的某些类型的细胞。这些发现表明，组蛋白H2B-GFP转基因的CMV驱动表达取决于基因组中转基因的整合位点。然而，通过使用转基因花aka，容易而精确地监测大多数有丝分裂细胞中核和染色体的细胞学变化，将极大地有助于更好地了解脊椎动物细胞中核和染色体行为的控制机制。

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