GENETIC AND ANATOMICAL MAP OF EXPRESSED SEQUENCES

我们建议建立一个关于 10,000-20,000个最常见的小鼠脑内mRNAs在 大脑中不同的解剖位置，在几个发育阶段 阶段，在外周组织中，并对生理和 行为范式。从数据中，将由一个新开发的 基于聚合酶链式反应的方法在概念上类似于但比 差异显示，我们将识别几千个差异 表达或生理调节的mRNAs。我们将确定部分 900个差异表达或调控的mRNAs序列，这将 为每个人提供身份标签，并导致小说的识别 蛋白质。将使用特定的聚合酶链式反应引物来建立表达 更高精密度的图谱和SSCP-PCR扩增 来自94只种间回交小鼠的基因组DNA 确定每个聚合酶链式反应产物在几cM范围内与已有标记的连锁 映射到面板上。从这些研究中将产生一个有注解的公众 数据库编目的聚合酶链式反应引物，聚合酶链式反应产物长度，相对数量 不同组织中的产物、部分核苷酸序列和基因 通过差异表达识别基因的位置 模式可以由科学家用特定的 用于深入研究的发育/生理/行为问题。 同样重要的是，遗传和表达模式方面将允许 中枢神经系统和遗传性疾病的候选基因 某些需要识别的外周组织。研究将会回答 基因数量和基因优先聚类问题 在特定的解剖和/或发育窗口转录，以及 提供了对基因表达模式的更好理解，从而 将为一项技术提供基线数据， 由于其对总基因表达的数字方法，一个主要的分析 辨别生理功能正常和缺陷的工具。

We propose to build a database of quantitative information concerning the expression of the 10,000 to 20,000 most prevalent mouse brain mRNAs in different anatomical sites within the brain, at several developmental stages, in peripheral tissues, and in response to physiological and behavioral paradigms. From the data, to be collected by a newly developed PCR-based methodology conceptually similar to but more exhaustive than differential display, we will recognize a few thousand differentially expressed or physiologically regulated mRNAs. We will determine partial sequences of 900 differentially expressed or regulated mRNAs, which will provide an identity tag for each and lead to the identification of novel proteins. Specific PCR primers will be used to establish expression profiles at higher degrees of precision and for SSCP-PCR amplification of genomic DNA from a panel of 94 interspecific backcross mice so as to determine linkage of each PCR product, within a few cM, to markers already mapped on the panel. From these studies will emerge an annotated public database cataloguing PCR primer, PCR product length, relative amounts of the products in different tissues, partial nucleotide sequence and gene location in which genes identified by their differential expression patterns can be identified by scientists with particular developmental/physiological/ behavioral questions for in depth studies. Equally important, the genetic and expression pattern aspects will allow candidate genes for genetically determined disorders of the CNS and certain peripheral tissues to be recognized. The studies will answer questions of gene number and clustering of genes preferentially transcribed in specific anatomical and/or developmental windows, and provide a better understanding of patterns of gene expression, and thus will provide baseline data for a technology that is certain to become, because of its digital approach to total gene expression, a major analytic tool for discerning normal and defective physiological function.