MAPPING NETWORKS OF GENE INTERACTIONS IN YEAST

绘制酵母基因相互作用网络图

• 批准号: 2611448
• 负责人: RONALD A BUTOW
• 金额: $ 46.75万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-08-15 至 2000-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2611448
• 关键词: Saccharomyces cerevisiae; biological transport; biomedical equipment; biomedical equipment development; fungal genetics; gene expression; gene interaction; genetic mapping; microorganism culture; nucleic acid hybridization

The overall objective of the proposed research is to identify and map networks of gene interactions in Saccharomyces cerevisiae. The strategy is to identify genes whose mRNA abundance is altered as a consequence of directed gene perturbations; initially, these will be single-gene replacements. The basic method to be used is the hybridization of cDNAs derived from wild-type and experimental strains to arrays of all known and predicted yeast genes. The hybridization signals will be quantified to obtain values of relative gene expression among the different experimental samples. As a model system, we will analyze gene interactions in a complex, poorly understood, interorganelle communication network called the retrograde response, in which changes in the functional state of mitochondria result in changes in expression of a large but unidentified subset of nuclear genes. Four specific aims are proposed: 1) to construct a high throughput microdisplay system, patterned after an existing design, for the analysis of expression of all known and predicted yeast genes; 2) to identify genes and pathways that constitute the retrograde response; 3) to develop general methodology for the identification, analysis and presentation of networks of gene interactions; and 4) to make the technology, reagents and data available to the scientific community. The general methodology will involve tabulation and graphical analysis of the data to obtain, as a long range goal, i) the relative expression value of any given gene in the yeast genome as it samples all viable single-gene replacements, and, as a short term goal, ii) an array of genes organized as nested sets whose expression changes in response to a given single-gene replacement, beginning with known genes in the retrograde response pathways. These studies are aimed at furthering our understanding of polygenic interactions that underlie poorly understood genetic phenomena such as penetrance and quantitative traits.

拟议研究的总体目标是确定和绘制 酿酒酵母中的基因相互作用网络。战略 是鉴定那些mRNA丰度因此而改变的基因 定向基因扰动;最初，这些将是单基因 替代品所用的基本方法是cDNA的杂交 从野生型和实验菌株衍生到所有已知的 并预测了酵母基因。将对杂交信号进行定量 为了获得不同基因组之间的相对基因表达值， 实验样品作为一个模型系统，我们将分析基因 在一个复杂的，知之甚少的， 一种称为逆行反应的通讯网络， 在线粒体的功能状态导致表达的变化 一个庞大但未被确认的核基因子集四个具体目标 提出：1）构建高吞吐量微显示系统， 根据现有的设计进行模式化，用于分析 所有已知和预测的酵母基因; 2）识别基因和途径 这构成了逆行反应; 3）发展一般 确定、分析和列报 基因相互作用的网络;以及4）为了使技术，试剂 和数据提供给科学界。的一般方法 将涉及数据的制表和图形分析， 作为一个长期目标，i）任何给定的相对表达值 基因在酵母基因组中，因为它样本所有可行的单基因 替代品，以及作为短期目标，ii）一系列基因 组织为嵌套集，其表达式会响应 给定单基因替换，从已知基因开始， 逆行反应途径这些研究旨在进一步促进我们的 对多基因相互作用的理解， 遗传现象，如遗传和数量性状。