HYBRIDOMAS: PRODUCTION AND GENETIC APPLICATIONS

杂交瘤：生产和遗传应用

• 批准号: 3166387
• 负责人: ROGER H KENNETT
• 金额: $ 11.28万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 1978
• 资助国家: 美国
• 起止时间: 1978-09-01 至 1989-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3166387
• 关键词: DNA; DNA binding protein; Escherichia coli; affinity chromatography; antibody specificity; antigens; binding proteins; biotechnology; cell differentiation; cell transformation; chemical structure function; cross immunity; electroporation; enzyme linked immunosorbent assay; fibroblasts; gene expression; genetic manipulation; genetic recombination; genetic transcription; hybridomas; immunoelectrophoresis; intermolecular interaction; laboratory mouse; microinjections; monoclonal antibody; neoplastic cell; nucleoproteins; oncogenes; oncoproteins; phosphoproteins; point mutation; protein sequence; radiotracer; site directed mutagenesis; transfection; tumor antigens

The goal of this work is to develop a detailed understanding of the structure and function of the c-myc gene product. The working hypothesis is that the c-myc protein is involved in the control of transcription and functions as part of a complex of proteins which bind to DNA. The intention is to use a combination of monoclonal antibody and recombinant DNA techniques to analyze the structure and function of this oncogene protein. A panel of epitope specific antibodies will be produced and characterized. These antibodies will be used to define epitopes on the surface of the protein, and regions of the molecule involved in interactions with DNA and with other proteins. Coprecipitation will be used in an attempt to detect proteins associated with c-myc. The antibodies will also be used to determine if there is structural homology between the c-myc protein and other proteins particularly those with known functions or others that bind DNA. In parallel with this work an assay for the biological activity of the c-myc will be developed based on either the transformation of primary lymphocytes or fibroblasts. This assay will then be used to evaluate the effects of site-directed mutagenesis on specific c- myc epitopes. The epitope specific antibodies will be assayed for their effects on the growth of cells into which they are incorporated by microinjection or electroporation. A long term goal is to understand the detailed structure and function of the c- myc protein. This may possibly lead to the ability to specifically modulate the activity of this protein that apparently plays an important role in the control of cell proliferation.

这项工作的目的是对 C-MYC基因产物的结构和功能。 工作 假设是C-MYC蛋白参与了 转录和功能是蛋白质复合物的一部分 与DNA结合。 目的是结合单克隆 抗体和重组DNA技术分析 该癌基因蛋白的结构和功能。 一个面板 将产生和表征表位特异性抗体。 这些抗体将用于定义表面 蛋白质和相互作用的分子区域 与DNA和其他蛋白质。 共沉淀将用于 试图检测与C-MYC相关的蛋白质。 这 抗体也将用于确定是否存在结构 C-MYC蛋白与其他蛋白质之间的同源性 特别是那些具有已知功能的人或其他结合DNA的人。 与这项工作并联 C-MYC将根据一个转换开发 原发性淋巴细胞或成纤维细胞。 然后将使用此测定 评估位置定向诱变对特定c-的影响 MYC表位。 表位特异性抗体将被分析 它们对细胞生长的影响 通过显微注射或电穿孔合并。 长期 目标是了解C-的详细结构和功能 MYC蛋白。 这可能会导致具体的能力 调节这种蛋白质的活性，显然是在 在控制细胞增殖中的重要作用。