DISCOVERY OF CSNPS IN NICOTINIC ACETYLCHOLINE RECEPTORS

烟碱乙酰胆碱受体中 CSNPS 的发现

• 批准号: 6187363
• 负责人: CARL C TON
• 金额: $ 25.58万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-09-30 至 2001-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6187363
• 关键词: RNA splicing; computer assisted sequence analysis; genetic polymorphism; genetic promoter element; human genetic material tag; human tissue; introns; neurogenetics; nicotinic receptors; nucleic acid sequence; polymerase chain reaction

DESCRIPTION: (Applicant's abstract) The goal of this pilot-scale project is to exhaustively discover and score for coding single nucleotide polymorphisms (cSNPs) within the family of genes that encode subunits of the neuronal nicotinic acetylcholine receptor (nAChR). The family comprises eleven known members: eight alpha-type subunits (alpha2 - alpha9) and three beta subunits (beta2 - beta4). Members of this class of pentameric ligand-gated Ca++ channels are known or suspected of being involved in a wide range of human disorders ranging from epilepsy and schizophrenia to nicotine addiction and auditory dysfunction. Thus a comprehensive set of cSNPs for these genes would greatly assist in future genetic and/or biological studies on human disease, or of gene/environment interactions. The specific aims of the project are: (1) gene structure: determination of the genomic organization (e.g. putative promoter region, intron/exon boundaries) of the 11 receptor genes, hence to develop robust PCR reactions for the amplification of all the exons and their flanking splice sites, as well as promoter regions. (2) Discovery of cSNPs: this will be accomplished through fluorescence labeled dye-terminator and gel-based resequencing of the PCR amplified exons. The PCR amplified exons will be derived from the DNA of 400 individuals drawn from the panel designated as the Resource for the Discovery of SNPs. To maximize the efficiency of single-pass sequencing in SNP detection, the base-calling program PolyPhred will be used to assist in the identification of heterozygous polymorphic sites. This measure will also allow a degree of automation and quality assessment of the SNP detection process at the level of each sequence read.

描述：（申请人摘要）本试验规模项目的目标是 为了彻底发现和评分编码单核苷酸 基因多态性（cSNP）的基因家族内的编码亚基的基因， 神经元烟碱乙酰胆碱受体（nAChR）。 家庭成员包括 11个已知成员：8个α型亚基（α 2-α 9）和3个 β亚基（β 2-β 4）。 五聚体的这一类别的成员 配体门控的Ca++通道是已知的或怀疑参与了一个 从癫痫和精神分裂症到 尼古丁成瘾和听觉功能障碍。 因此， 这些基因的cSNPs将极大地有助于未来的遗传和/或 对人类疾病或基因/环境相互作用的生物学研究。 该项目的具体目标是：（1）基因结构：确定 基因组组织（例如推定的启动子区、内含子/外显子 边界）的11个受体基因，从而开发强大的PCR反应 用于扩增所有外显子及其侧翼剪接位点，如 以及启动子区。 (2)cSNPs的发现：这将是 通过荧光标记的染料终止剂和凝胶实现 PCR扩增的外显子的重测序。 PCR扩增的外显子将被 来自400个个体的DNA，这些个体来自指定为 SNPs发现资源。 效率最大化 单次测序在SNP检测中，碱基调用程序PolyPhred 将用于协助鉴定杂合多态性 网站. 这项措施也将允许一定程度的自动化和质量 在每个序列读段的水平上评估SNP检测过程。