HUMAN COCHLEAR CDNA CLONES

这项计划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表征从胎儿耳蜗cDNA文库中识别的那些cDNA克隆来识别，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着的序列同源性。将通过筛选耳蜗CAPfinder文库获得全长cDNA序列，并通过BLAST分析进行仔细检查。将评估表达模式未知的cDNA克隆的RNA表达模式。将通过细胞遗传学分析和辐射杂交作图评估染色体图谱位置。任何发现映射到耳聋基因座的cDNA都可以在相关患者中筛选突变。那些发现是有趣的cDNA（即，耳聋、基因座、耳蜗表达的图谱）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进一步表征以确定蛋白质定位。

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.