ANALYSIS OF REGULATION OF MUCIN GENE EXPRESSION

(根据调查员的具体目的改编)这项工作的具体目的 修改后的应用是：1)从大鼠和大鼠中分离全长粘蛋白cDNA 人类气道库；从这些库中确定了 两个物种的粘蛋白核心蛋白。2)利用大鼠粘蛋白基因，分离 从大鼠基因组文库中克隆同源基因并鉴定上游(或 其他)利用缺失的转染法分析调控元件 变种人。3)确定这些监管要素的约束程度 使用迁移率变化和DNase I保护分析的核蛋白和 4)建立大鼠呼吸道粘蛋白过度产生模型，并测定 粘蛋白生物合成的转录控制是否上调 大鼠粘蛋白cdna的Northern杂交和核连续试验。

(Adapted from the investigator's specific aims) The specific aims of this revised application are: 1) Isolate full length mucin cDNAs from rat and human airway libraries; from these determine the primary structure of mucin core protein in both species. 2) Using the rat mucin cDNA, isolate the cognate gene from a rat genomic library and identify upstream (or other) regulatory elements using transfection analysis of deletion mutants. 3) Determine the extent to which these regulatory elements bind nuclear proteins using mobility shift and dNase I protection assays and 4) Establish a rat model of respiratory mucin overproduction and determine whether transcriptional control of mucin biosynthesis is upregulated using a rat mucin cDNA in northern hybridization and nuclear run-on assays.