GENETIC BASIS OF ANTIGENIC VARIATION

抗原变异的遗传基础

• 批准号: 3481658
• 负责人: ROBERT A HYMAN
• 金额: $ 31.45万
• 依托单位: SALK INSTITUTE FOR BIOLOGICAL STUDIES
• 依托单位国家: 美国
• 财政年份: 1978
• 资助国家: 美国
• 起止时间: 1978-06-01 至 1993-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3481658
• 关键词: DNA binding protein; Retroviridae; cell fusion; gene expression; genetic manipulation; genetic transcription; histocompatibility antigens; histocompatibility gene; hybrid cells; immunogenetics; immunoglobulin genes; laboratory mouse; laboratory rabbit; lymphoma; neoplasm /cancer genetics; nuclear runoff assay; southern blotting; surface antigens; tissue /cell culture; transcription factor; transfection; transposon /insertion element; tumor antigens

Cell surface molecules that are differentially expressed on particular subpopulations of cells may act to regulate cell growth and differentiation or mediate cell interactions with the external environment. Transformed cells may show abnormal expression of particular cell surface molecules, and, in some cases, this abnormal expression may mediate the transformed state. The objective of this project is to define genes that control the cell surface expression of the Thy-1 and Lyt-2 glycoproteins and to determine their mechanism of action. Specific somatic cell mutants provide a way to study the control of the expression of cell surface molecules, particularly when coupled with the use of molecular genetic techniques to introduce specific genes into mutant cell lines. A Thy-1- mutant that acts in trans position to extinguish expression of Thy-1 in hybrids of the mutant with wild-type Thy-1+ cell lines has been described. Nuclear runoff transcription assays will be used to determine whether the gene product defined by this mutant acts transcriptionally. Proteins that bind to the DNA of this Thy-1- mutant and its Thy-1+ revertant will be characterized. Transfection and somatic cell fusion will be used to define regions of the Thy-1 gene that interact with the gene product defined by this mutant. Retroviral insertional mutagenesis will be used to attempt to identify and isolate the gene defined by the mutant. Somatic cell mutants will also be used to define regions important in controlling the expression of the Lyt-2 gene. Southern blotting analysis of one mutant will be used to define a sequence that acts in cis position to activate Lyt-2 expression The gene(s) that act to regulate Lyt-2 and Thy-1 expression in interlineage hybrids will be characterized using a combination of segregation analysis and the introduction of genes into hybrids by transfection.

细胞表面差异表达分子 特定的细胞亚群可能起到调节细胞生长的作用 和分化或介导细胞与外界的相互作用 环境。 转化细胞可能表现出异常表达 特定的细胞表面分子，并且在某些情况下，这 异常表达可能介导转化状态。 这 该项目的目标是定义控制细胞的基因 Thy-1 和 Lyt-2 糖蛋白的表面表达 确定它们的作用机制。 特定的体细胞突变体提供了一种研究控制的方法 细胞表面分子的表达，特别是当 再加上使用分子遗传学技术来引入 将特定基因导入突变细胞系。 一种 Thy-1- 突变体，作用于 反位以消除 Thy-1 在杂合体中的表达 野生型 Thy-1+ 细胞系的突变体已被描述。 核径流转录测定将用于确定 该突变体定义的基因产物是否起作用 转录地。 与 Thy-1- DNA 结合的蛋白质 突变体及其 Thy-1+ 回复体将被表征。 转染和体细胞融合将用于定义区域 Thy-1 基因与以下定义的基因产物相互作用 这个突变体。 逆转录病毒插入诱变将用于 尝试识别和分离突变体定义的基因。 体细胞突变体也将用于定义重要区域 控制 Lyt-2 基因的表达。 南方印迹 对一个突变体的分析将用于定义发挥作用的序列 处于顺式位置以激活 Lyt-2 表达 调节 Lyt-2 和 Thy-1 表达的基因 系间杂种将使用以下组合来表征 分离分析和将基因引入杂种 转染。