HUMAN COCHLEAR CDNA CLONES

人类耳蜗 CDNA 克隆

• 批准号: 6329201
• 负责人: BARBARA L RESENDES
• 金额: $ 4.02万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-12-01 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6329201
• 关键词: chromosomes; cochlea; complementary DNA; gene expression; genetic library; hearing; messenger RNA; molecular cloning; molecular genetics; protein localization

The long-term objective of this proposal is to identify human auditory genes and determine their role in the auditory process. The human auditory genes will be identified by characterizing those cDNA clones identified from a fetal cochlear cDNA library that had preferential cochlear mRNA expression patterns or significant sequence homology to non-human genes thought to be important for the hearing process. Full-length cDNA sequences will be obtained by screening a cochlear CAPfinder library and scrutinized by BLAST analysis. RNA expression patterns will be assessed for those cDNA clones whose expression patterns are not known. The chromosomal map position will be assessed by cytogenetic analysis and radiation hybrid mapping. Any cDNA found to map to a deafness loci may be screened for mutations in relevant patients. Those cDNAs found to be interesting (i.e., maps to deafness, loci, cochlea expression) will be characterized further by in situ hybridization for determining the cell type expression patterns and by immunohistochemistry to determine protein localization.

这项提议的长期目标是确定人类听觉基因并确定它们在听觉过程中的作用。人类听觉基因将通过鉴定从胎儿耳蜗cDNA文库中鉴定的cDNA克隆来鉴定，这些克隆具有优先的耳蜗mRNA表达模式或与被认为对听力过程重要的非人类基因具有显著的序列同源性。通过筛选耳蜗CAPfinder文库获得全长cDNA序列，并进行BLAST分析。RNA表达模式将评估那些表达模式未知的cDNA克隆。染色体图谱位置将通过细胞遗传学分析和辐射杂交作图来评估。任何发现与耳聋基因位点相关的cDNA都可以在相关患者中进行突变筛选。那些发现有趣的cdna（即耳聋、基因座、耳蜗表达图）将通过原位杂交进一步表征，以确定细胞类型表达模式，并通过免疫组织化学确定蛋白质定位。