MOLECULAR AND GENETIC ANALYSIS OF ALBUMIN EXTINCTION

白蛋白消失的分子和遗传分析

• 批准号: 3087075
• 负责人: AMELIA A LANGSTON
• 金额: $ 8.82万
• 依托单位: FRED HUTCHINSON CANCER RESEARCH CENTER
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-08-01 至 1995-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3087075
• 关键词: DNA; albumins; alcohol dehydrogenase; cell fusion; developmental genetics; electrophoresis; gene expression; gene induction /repression; genetic manipulation; hybrid cells; karyotype; molecular cloning; nucleic acid hybridization; transcription factor

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.

我们正在使用体细胞和分子遗传学技术来研究基因座 调控基因的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 通常不表达亲本中任何一方的组织特异性产物， 这一现象被称为灭绝。两个离散的遗传基因座Tse-1和Tse-2， 已经被证明是导致不同肝脏的灭绝的原因- 肝癌-成纤维细胞杂交瘤中的特异性基因。这是一项旨在 对其中一个基因座Tse-2进行详细研究。 这个项目的目的是确定TSE-2调解的手段 一组肝脏基因的消失，包括白蛋白和酒精 脱氢酶。TSE-2已经被定位到小鼠的1号染色体上，尽管 稳定的单染色体杂交种不能用于研究 TSE-2。该项目的初始阶段将使用微细胞介导的 染色体转移产生稳定的单染色体杂交种 一个活跃的Tse-2基因座。经核型分析和Southern杂交分析 灭活的克隆，它们将被用于研究Tse-2+在 详细信息，并调查TSE-2与其他已知的 受影响基因的转录调节因子。这些混合动力车将 也可作为消减克隆的起始材料。 TSE-2的文字记录。