MAPPING NETWORKS OF GENE INTERACTIONS IN YEAST

拟议研究的总体目标是确定和绘制地图 酿酒酵母中的基因互作网络。战略 就是找出基因的信使核糖核酸丰度因此改变 定向基因干扰；最初，这些将是单基因 接班人。要使用的基本方法是cDNA杂交 从野生型和实验菌株衍生到所有已知的阵列 并预测了酵母基因。杂交信号将被量化 为了获得不同物种之间的相对基因表达值 实验样本。作为一个模型系统，我们将分析基因 复杂的、鲜为人知的细胞器间的相互作用 通讯网络称为逆行反应，在其中发生变化 线粒体的功能状态导致表达的变化 一个庞大但未被识别的核基因子集。四个具体目标 提出：1)构建高通量微显示系统， 仿照现有设计，用于分析 所有已知和预测的酵母基因；2)确定基因和途径 构成逆行反应；3)发展一般反应 确定、分析和介绍 基因相互作用网络；以及4)使这项技术、试剂 以及科学界可用的数据。一般方法论 将涉及数据的制表和图形分析以获得， 作为一个长期目标，i)任何给定的相对表达值 酵母基因组中的基因，因为它采样了所有活的单基因 替代，以及，作为短期目标，ii)一系列基因 组织为嵌套集，其表达式随 给予单基因替换，从已知的基因开始 逆行反应通路。这些研究的目的是进一步推动我们的 对多基因相互作用的理解知之甚少 遗传现象，如外显性和数量性状。

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.