GENETIC AND ANATOMICAL MAP OF EXPRESSED SEQUENCES

表达序列的遗传和解剖图

• 批准号: 2272193
• 负责人: J GREGOR SUTCLIFFE
• 金额: $ 47.95万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-04-01 至 1998-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2272193
• 关键词: animal genetic material tag; brain mapping; computer assisted sequence analysis; developmental genetics; gene expression; genetic mapping; laboratory mouse; linkage mapping; polymerase chain reaction

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.

我们建议建立一个关于 表达的10，000至20，000最普遍的小鼠脑mRNAs中， 大脑中不同的解剖部位，在几个发育阶段， 阶段，在外周组织中，并响应于生理和 行为模式 从新开发的数据中收集 基于PCR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mRNA。 我们将确定部分 900个差异表达或调节的mRNA序列， 为每个人提供身份标签并识别小说 proteins. 将使用特异性PCR引物建立表达 在更高的精确度和SSCP-PCR扩增的概况 一组94只种间回交小鼠的基因组DNA， 确定每个PCR产物的连锁，在几厘米内，标记已经 映射在面板上。 从这些研究中将出现一个注释的公众 数据库编目PCR引物，PCR产物长度， 不同组织中的产物、部分核苷酸序列和基因 通过差异表达鉴定基因的位置 科学家可以通过特定的 发展/生理/行为问题进行深入研究。 同样重要的是，基因和表达模式方面将允许 用于遗传决定的CNS病症的候选基因， 某些外围组织被识别。 研究将回答 基因数和基因优先聚类问题 在特定的解剖学和/或发育窗口中转录，以及 更好地了解基因表达模式，从而 将为这项技术提供基准数据，这项技术肯定会成为， 由于其对总基因表达的数字化方法， 用于辨别正常和有缺陷的生理功能的工具。