MOLECULAR AND GENETIC ANALYSIS OF ALBUMIN EXTINCTION

我们正在使用体细胞和分子遗传技术来研究基因座 调节基因发育和组织特异性模式 表情 不同类型细胞融合形成的体细胞杂种 通常不能表达任一亲本的组织特异性产物， 这种现象称为灭绝。 两个离散的遗传位点Tse-1和Tse-2， 已经被证明会导致不同类型的肝脏灭绝 肝癌-成纤维细胞杂种中的特定基因。 这是一个建议， 研究这些位点之一，Tse-2，详细。 本项目的目的是确定Tse-2介导的手段 一组肝脏基因的灭绝，包括白蛋白和酒精 脱氢酶。 TSE-2已定位于小鼠1号染色体， 稳定的单染色体杂种不能用于促进 Tse-2。 该项目的初始阶段将使用微细胞介导的 染色体转移以产生稳定的单染色体杂种，所述单染色体杂种包含 一个活跃的Tse-2位点 经核型和Southern印迹分析， 灭绝的克隆，它们将用于研究Tse-2+表型， 详细，并调查Tse-2和其他已知的 受影响基因转录的调节因子。 这些混血儿 也可用作cDNA消减克隆的起始材料， TSE-2转录本。

We are using somatic cell and molecular genetic techniques to study loci that regulate developmental and tissue-specific patterns of gene expression. Somatic hybrids formed by fusing cells of different types generally fail to express the tissue specific products of either parent, a phenomenon termed extinction. Two discrete genetic loci, Tse-1 and Tse-2, have been shown to contribute to the extinction of distinct sets of liver- specific genes in hepatoma-fibroblast hybrids. This is a proposal to investigate one of these loci, Tse-2, in detail. The aim of this project is to determine the means by which Tse-2 mediates extinction of a group of liver genes, including albumin and alcohol dehydrogenase. Tse-2 has been mapped to murine chromosome 1, although stable monochromosomal hybrids are not available to facilitate study of Tse-2. The initial phase of this project will use microcell-mediated chromosome transfer to generate stable monochromosomal hybrids containing an active Tse-2 locus. After karyotypic and Southern blot analysis of extinguished clones, they will be used to study the Tse-2+ phenotype in detail, and to investigate the relationship between Tse-2 and other known regulators of transcription of the affected genes. These hybrids will also be used as starting materials for cDNA subtractive cloning of the Tse-2 transcript.