MOLECULAR CLONING OF C-MYC TRANSCRIPTION FACTOR GENES

C-MYC 转录因子基因的分子克隆

• 批准号: 3432472
• 负责人: PAUL Eric NEIMAN
• 金额: $ 2.08万
• 依托单位: FRED HUTCHINSON CANCER RESEARCH CENTER
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-02-20 至 1995-02-19
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3432472
• 关键词: B lymphocyte; DNA binding protein; DNA footprinting; bursa of Fabricius; chickens; complementary DNA; genetic regulation; genetic regulatory element; genetic transcription; molecular cloning; protooncogene; tissue /cell culture; transcription factor; transfection

Dr. Victor Lobanenkov, is experienced in identifying and characterizing trans-acting transcription factors. He plans to collaborate with Dr. Paul Neiman to exploit the biological systems Dr. Neiman has characterized, namely the development and differentiation of avian B-cells in the bursa of Fabricius in which the controlled expression of the c-myc proto-oncogene plays an important role. We plan to identify the regulatory factors which are involved in regulating initiation and/or elongation of the transcription of the c-myc gene with the major emphasis toward understanding how these control the gene in bursal B-cells. Having identified the DNA-binding proteins involved, the cDNAs encoding these proteins will be isolated. These cDNAs will be used to study expression of the genes in B-cells and retrovirus vectors will be used to reintroduce the' genes into B-cells, either as constitutively active factors or as "dominant negative" factors, to investigate their effects on B-cell development. The cloned genes will also be used to express the proteins in E. coli for studies of protein interaction. As part of the research program, we wish to identify cis and trans elements involved in myc negative autoregulation. To this end, we plan to use a recently developed approach based on retrovirus construct transducing both expression of the conditional myc-fusion protein and a selectable marker.

维克托·洛巴年科夫博士，在鉴别和描述 反式作用转录因子。 他计划与博士合作。 保罗·尼曼利用尼曼博士的生物系统 特征，即鸟类的发育和分化 在法氏囊中的B细胞中， c-myc原癌基因起重要作用。 我们计划找出 参与调节起始和/或 延长c-myc基因的转录，主要强调 以了解它们如何控制囊B细胞中的基因。 在鉴定了所涉及的DNA结合蛋白之后， 这些蛋白质将被分离。 这些cDNA将用于研究 在B细胞和逆转录病毒载体中的基因表达将用于 将基因重新引入B细胞，无论是作为组成型活性， 因素或“显性负面”因素，以调查其影响 对B细胞发育的影响 克隆的基因也将用于表达 E. coli中进行蛋白质相互作用的研究。 的一部分 研究计划，我们希望确定涉及顺式和反式元素， myc负性自身调节。 为此，我们计划最近使用一个 基于逆转录病毒构建体开发的方法， 条件性myc融合蛋白和选择标记的表达。