GENE EXPRESSION DURING ERYTHROID MATURATION

我们的长期目标是从免疫学上揭示 细胞阶段特异性变化的表面蛋白与 小鼠红系集落形成单位（CFU-E）的成熟， 为了分离编码这些蛋白质的重组克隆， cDNA表达文库，并使用这样的克隆来评估阶段- 特定基因水平的事件。 我们的假设是红细胞 成熟提供了一个结束阶段的情况下， 细胞阶段的具体事件发生，但当设置对 生物体发育中遇到的变化幅度， 在数量上相对适中。 在这个可能简化的 设置，我们的目的是建立一个经常匿名的剧目 但是我们和其他人可以用差异表达的基因 我希望开始剖析整个监管事件 红细胞成熟的某些方面。 七项具体 目标是：首先扩大我们现有的老鼠收集 单克隆抗体（抗鼠细胞表面抗原的单克隆抗体 在红细胞的特定阶段优先表达 成熟;第二次鉴定特别适合以后使用的MoAb; 第三，补充我们现有的网织红细胞缺乏球蛋白的cDNA 表达式库与加/减一;第四，通过应用 筛选单克隆抗体，对表达文库进行筛选， 编码细胞表面优先表达的cDNA克隆 元素;第五，验证特定于阶段的表达式， 第六，分离相应的基因组DNA序列; 第七，利用cDNA克隆和相应的 （a）基因组序列，以表征特定基因结构， 与差异基因表达相关的基因水平事件 以及（B）最终，鉴定这些基因座 及其调节基因表达变化的产物 在我们的剧目。

Our long-range goals are to uncover immunologically-a portion of the cell stagespecific changes in surface proteins associated with the maturation of murine erythroid colony-forming units (CFU-E), to isolate recombinant clones encoding some of these proteins from cDNA-expression libraries, and to use such clones to assess stage- specific gene-level events. Our assumption is that erythroid maturation provides an end-stage situation in which a substantial body of cell-stage specific events occur but, when set against the magnitude of change encountered in organismal development, are comparatively modest in number. In this potentially simplified setting, our intent is to establish a repertoire of often anonymous but differentially expressed genes with which we and others can hope to begin dissecting the ensemble of regulatory events underlying some aspects of erythroid maturation. Seven Specific Aims are set: first enlarge our existing collection of rat monoclonal antibodies (MoAb against murine cell-surface antigens preferentially expressed at a particular stage of erythroid maturation; second identify MoAb especially suitable for later use; third, supplement our existing reticulocyte globin-poor-cDNA expression library with a plus/minus one; fourth, by applying selected MoAb to the screening of expression libraries, isolate cDNA clones encoding preferentially expressed cell-surface elements; fifth, validate stage-specific expression of thus isolated cDNA; sixth, isolate genomic DNA sequences corresponding to such cDNA; and seventh, utilize cDNA clones and corresponding genomic sequences (a) to characterize specific gene structures and gene-level events associated with differential gene expression during erythroid maturation and (b) ultimately, identify those loci and their products which modulate changing expression of the genes in our repertoire.