SGER: Development of GFP-chromosome Tagging System

酿酒酵母Dawson 9610330因其遗传易感性而成为研究减数分裂染色体行为的理想生物。作为研究减数分裂的生物，它的主要缺点是染色体很小，在减数分裂细胞中观察不到它们的行为。这项用于探索性研究的小额赠款的目标是开发一种新的系统，可以在活的减数分裂酵母细胞中可视化染色体。该系统将基于绿色荧光蛋白杂交分子与放置在特定染色体上的位点的结合。目前对减数分裂染色体行为的了解主要是通过遗传学和细胞生物学研究相结合的方式收集的。通常，这些研究是在不同的生物体中进行的。例如，酵母很好地用于遗传研究，而植物和昆虫，因为它们的染色体可以用光学显微镜在活细胞中可视化，为细胞生物学家提供了宝贵的见解。这些方法的结合将特别强大，因为它将使遗传分析已知在减数分裂染色体分离(配对、重组、联会复合体形成、纺锤体组装和染色体运动)方面有缺陷的突变体的染色体行为可视化。在过去的十年中，发现了大量具有减数分裂缺陷的酵母突变株。在这些突变体和野生型细胞中观察染色体行为的系统的发展将对我们理解建立适当的染色体行为的方式产生巨大影响，这是一个对所有有性繁殖高等生物至关重要的过程。

Dawson 9610330 The yeast Saccharomyces cerevisiae has been a favored organism for the study of meiotic chromosome behavior because of its genetic tractability. Its primary deficiency as an organism for studying meiosis is that its chromosomes are very small and their behavior can not be observed in meiotic cells. The goal of this Small Grant for Exploratory Research is to develop a novel system that could allow the visualization chromosomes in living meiotic yeast cells. The system would be based on the binding of green fluorescent protein hybrid molecules to sites placed on specific chromosomes. Current understanding of the behavior of meiotic chromosomes has been gleaned primarily by a combination of genetic and cell biological studies. Often these studies have been performed in different organisms. For example, yeast has lent itself well to genetic studies while plants and insects, because their chromosomes can be visualized in living cells with a light microscope, have provided cell biologists with invaluable insights. The coupling of these approaches would be especially powerful because it would permit the visualization of chromosome behavior in mutants known by genetic analysis to be defective in aspects of meiotic chromosome segregation (pairing, recombination, synaptonemal complex formation, spindle assembly and chromosome movement). In the past decade a large collection of yeast mutants with meiotic defects have been identified. The development of a system for viewing chromosome behavior in these mutants and wild type cells would have a tremendous impact on our understanding of the manner in which proper chromosome behavior is established, a process essential to all sexually reproducing higher organisms.