MOLECULAR CLONING OF C-MYC TRANSCRIPTION FACTOR GENES

维克托·洛巴年科夫博士在识别和描述 反式作用转录因子。他计划与博尔顿博士合作。 保罗·内曼将利用内曼博士拥有的生物系统 特征，即鸟类的发育和分化 法氏囊中的B细胞，在其中控制表达 C-myc原癌基因在肿瘤发生发展中起重要作用。我们计划找出 参与调节启动和/或调节的调节因素 延长c-myc基因转录的主要侧重点 以了解这些基因如何控制法氏囊B细胞中的基因。 在确定了涉及的DNA结合蛋白后，编码 这些蛋白质将被分离出来。这些DNA将被用来研究 基因在B细胞和逆转录病毒载体中的表达将用于 将基因重新引入B细胞，无论是作为组成部分的活性 因素或作为“显性负面”因素，以调查其影响 关于B细胞的发育。克隆的基因也将被用来表达 用于研究蛋白质相互作用的大肠杆菌中的蛋白质。作为该计划的一部分 研究计划，我们希望确定参与的顺式和反式元件 MYC负性自动调节。为此，我们计划使用最近的一个 基于逆转录病毒构建转导两者的新方法 条件性myc融合蛋白和可选择标记的表达。

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.