酿酒酵母减数分裂染色体的实时成像
Real-time imaging of meiotic chromosomes in Saccharomyces cerevisiae.
作者信息
Koszul Romain, Kameoka Sei, Weiner Beth M
机构信息
Department of Molecular and Cellular Biology, Harvard University, Cambridge, MA, USA.
出版信息
Methods Mol Biol. 2009;558:81-9. doi: 10.1007/978-1-60761-103-5_6.
Important information on cellular physiology can be obtained by directly observing living cells. The nucleus, and the chromatin within, is of particular interest to many researchers. Monitoring the behavior of specific DNA loci in the living cell is now commonly achieved through the insertion of binding sites for fluorescently tagged proteins at the sequence of interest (e.g. Ref 1). However, visualizing the behavior of full length chromosomes can only be achieved when they constitute discrete, relatively well individualized units. During meiotic mid-prophase, chromosomes of budding yeast are well-organized structures that present such characteristics, making them remarkably suited for visualization. Here we describe the optimized protocols and techniques that allow monitoring of chromosome behavior during meiotic prophase in budding yeast.
通过直接观察活细胞可以获得有关细胞生理学的重要信息。细胞核以及其中的染色质是许多研究人员特别感兴趣的对象。目前，通过在感兴趣的序列处插入荧光标记蛋白的结合位点来监测活细胞中特定DNA位点的行为是很常见的做法（例如参考文献1）。然而，只有当全长染色体构成离散的、相对易于区分的单元时，才能观察到它们的行为。在减数分裂前期中期，出芽酵母的染色体是具有这些特征的组织良好的结构，这使得它们非常适合进行观察。在这里，我们描述了优化的方案和技术，这些方案和技术能够监测出芽酵母减数分裂前期的染色体行为。
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