GENE EXPRESSION DURING ERYTHROID MATURATION

我们的长期目标是从免疫学上发现 与细胞表面蛋白相关的细胞阶段性变化 小鼠红系集落形成单位(CFU-E)的成熟， 从这些蛋白质中分离编码部分蛋白质的重组克隆 表达文库，并利用这些克隆来评估阶段- 特定的基因水平事件。我们的假设是红血球 成熟提供了一种结束阶段的情况，在这种情况下， 特定于细胞阶段的事件的主体发生，但当设置为与 在生物体发育中遇到的巨大变化，是 在数量上相对较少。在这个可能被简化的 背景下，我们的目的是建立一个通常是匿名的曲目 但我们和其他人可以利用差异表达的基因 希望开始剖析一系列监管事件 在红系成熟的某些方面。七个具体的 目标已经确定：首先，扩大我们现有的老鼠集合 抗小鼠细胞表面抗原的单抗(单抗 在红系的特定阶段优先表达 成熟；第二，确定摩押特别适合以后使用； 第三，补充我们现有的网织红细胞珠蛋白贫乏基因 带正/负1的表达式库；第四，通过应用 选用单抗进行表达文库的筛选、分离 编码细胞表面优势表达基因的cDNA克隆 元素；第五，验证特定于阶段的表达式 分离的cDNA；第六，分离相应的基因组DNA序列 第七，利用cDNA克隆和相应的 基因组序列(A)以表征特定的基因结构和 与差异基因表达相关的基因水平事件 在红系成熟期间和(B)最终确定这些基因座 以及他们的调节基因表达变化的产物 在我们的剧目中。

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.