STRUCTURE AND EXPRESSION OF COMPLEMENT GENES C3 AND C4

补体基因C3和C4的结构和表达

• 批准号: 3129005
• 负责人: GEORG H FEY
• 金额: $ 15.99万
• 依托单位: SCRIPPS CLINIC AND RESEARCH FOUNDATION
• 依托单位国家: 美国
• 财政年份: 1983
• 资助国家: 美国
• 起止时间: 1983-01-01 至 1987-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3129005
• 关键词: alleles; chromosome complement; gel electrophoresis; gene expression; genetic manipulation; genetic mapping; genetic transcription; genome; hepatocellular carcinoma; human tissue; immune response genes; immunodeficiency; inflammation; macrophage; messenger RNA; molecular cloning; nucleic acid sequence; tissue /cell culture

The project proposes to study (a) the structure of the gene encoding complement protein C3, (b) the controls of transcription of this gene into RNA, (c) the molecular defects that lead to inherited C3 deficiencies in man and (d) the isolation of mouse cDNA. The study will take advantage of mouse C3 cDNA clones and C3 genomic DNA clones that have been prepared in the applicant's laboratory and will extend currently ongoing work. First, the tissue specific expression of C3 will be investigated. C3 mRNA concentrations will be titrated in hepatocytes, macrophages and in a variety of C3 producing and non-producing tissue culture cell lines and correlated with the state of methylation of the gene. The 5'-ends of liver and macrophage C3 mRNAs will be compared in order to determine whether the gene is transcribed in both tissues from the same promoter. It is intended to establish the total nucleotide sequence of C3 mRNA as a basis for future studies of particular regions of the C3 polypeptide, such as the binding sites for cellular C3d receptors. The second part proposes to isolate one intact copy of the human C3 gene from a gene library to be constructed in cosmid vectors. The human gene will be characterized and assayed for function by transfection into tissue culture cells. A defective allele carried by homozygous C3 deficient members of a Dutch family will be analyzed. In the third part an approach is described for the production of cloned mouse C4 cDNA clones. The last section outlines how C4 cDNA clones will be characterized and distinguished from cDNA clones for the C4 variant of mice, the Slp-protein. These clones will be used to screen mouse gene libraries for C4 genomic clones. Work on the human C4 gene will follow according to the initial results obtained with the mouse C4 gene. The proposed studies on the C3 and C4 genes will provide basic information on the structure and the regulation of these genes and also on inherited human disorders associated with these genes.

该项目建议研究（a）基因编码的结构 补体蛋白C3，（B）控制该基因转录成 RNA，（c）导致遗传性C3缺陷的分子缺陷， 人和（d）小鼠cDNA的分离。 这项研究将利用 小鼠C3 cDNA克隆和C3基因组DNA克隆， 申请人的实验室，并将扩大目前正在进行的工作。 第一、 将研究C3的组织特异性表达。 C3 mRNA 将在肝细胞、巨噬细胞和 多种C3生产和非生产组织培养细胞系， 与基因的甲基化状态相关。 肝脏5 ′端 和巨噬细胞C3 mRNA进行比较，以确定 基因在两种组织中从相同的启动子转录。 旨在 建立C3 mRNA的总核苷酸序列，作为未来研究的基础。 C3多肽的特定区域的研究，如结合 细胞C3d受体的位点。 第二部分提出隔离一个 从基因文库中构建人C3基因的完整拷贝， 粘粒载体。 将对人类基因进行表征和分析， 通过转染到组织培养细胞中发挥功能。 缺陷等位基因 携带纯合子C3缺陷成员的荷兰家庭将是 分析了 在第三部分中，描述了用于产生 克隆小鼠C4 cDNA克隆。 最后一节概述了C4 cDNA克隆 将被表征并与C4变体的cDNA克隆区分开 Slp蛋白。 这些克隆将用于筛选小鼠基因 C4基因组克隆的文库。 人类C4基因的研究工作将随之展开。 根据小鼠C4基因获得的初步结果， 的 对C3和C4基因的拟议研究将提供以下基本信息： 这些基因的结构和调控，以及遗传的人类 与这些基因相关的疾病。