HUMAN COCHLEAR CDNA CLONES

这项提议的长期目标是识别人类听觉基因，并确定它们在听觉过程中的作用。人类的听觉基因将通过鉴定那些从胎儿耳蜗库中鉴定出来的、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有显著序列同源性的cDNA克隆来识别。全长cDNA序列将通过筛选耳蜗CAPfinder文库获得，并通过BLAST分析进行筛选。对于那些表达模式未知的cdna克隆，将评估其RNA表达模式。染色体定位将通过细胞遗传学分析和辐射杂交作图进行评估。任何被发现映射到耳聋基因座的基因都可以在相关患者中进行突变筛查。那些被发现感兴趣的cDNA(即，与耳聋、基因、耳蜗表达的图谱)将进一步通过原位杂交来确定细胞类型的表达模式，并通过免疫组织化学来确定蛋白质的定位。

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.