MALE GERM CELL-AFFECTING GENES ON MOUSE CHROMOSOME 17

小鼠 17 号染色体上影响雄性生殖细胞的基因

• 批准号: 3326627
• 负责人: MASANORI KASAHARA
• 金额: $ 15.3万
• 依托单位: UNIVERSITY OF MIAMI SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-05-01 至 1992-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3326627
• 关键词: Escherichia coli; antibody; antisense nucleic acid; chromosomes; complementary DNA; fertility; gel electrophoresis; gene expression; gene mutation; genes; genetic library; genetic manipulation; genetic mapping; genetic transcription; genetically modified animals; genome; germ cells; laboratory mouse; laboratory rat; messenger RNA; molecular cloning; nucleic acid hybridization; proteins; spermatogenesis; testis

Numerous genes that affect spermatogenesis have been mapped to mouse chromosome 17. The overall goal of the proposed project is to isolate these genes by molecular cloning and to elucidate the mechanisms by which their mutations cause male sterility or sperm dysfunction. Since no information is available on their gene products, the following two cloning strategies will be used: (i) isolation of chromosome 17-derived genomic clones either at random or from the vicinity of HpaII tiny fragment (HTF) islands that exist in association with genes, and (ii) isolation of chromosome 17-encoded cDNA clones that are differentially expressed between normal and sterile mouse testes. The chromosome 17-derived genomic clones isolated from the vicinity of HTF islands will be screened for the presence of testicular transcripts, whereas the clones isolated at random will be regionally localized to identify those clones that map in the vicinity of known male germ cell-affecting loci. The clones that map close enough to such loci will be used to construct "regional" genomic libraries, which will then be screened for the presence of testicular transcripts. In our preliminary studies, the use of the first cloning strategy has resulted in the isolation of a novel testis-specific gene that apparently does not correspond to any of the known male germ cell- affecting loci located on chromosome 17. This gene, the protein products of which contain putative "zinc finger" motifs, will also be characterized in detail in this proposal. It is likely that the mouse genes we will isolate and characterize here have their human counterparts. Some forms of human male infertility may be accounted. for by their mutations. In this sense, the proposed project has direct clinical relevance and should provide us with basic knowledge that should lead to better diagnosis and treatment of human male infertility.

许多影响精子发生的基因已被定位到 小鼠染色体 17。该项目的总体目标是 通过分子克隆分离这些基因并阐明其作用 它们的突变导致男性不育或精子的机制 功能障碍。 由于没有关于其基因的信息 产品，将使用以下两种克隆策略：（i） 随机分离源自 17 号染色体的基因组克隆 或来自 HpaII 微小碎片 (HTF) 岛屿附近 与基因相关存在，并且（ii）染色体的分离 17 个编码的 cDNA 克隆在 正常且无菌的小鼠睾丸。 17号染色​​体衍生的基因组 将筛选从 HTF 岛屿附近分离出的克隆 睾丸转录本的存在，而克隆 随机隔离将进行区域本地化以识别那些 定位在已知影响雄性生殖细胞的附近的克隆 基因座。 将使用与这些基因座足够接近的克隆 构建“区域”基因组文库，然后将其 筛查睾丸转录本的存在。 在我们的 初步研究表明，第一个克隆策略的使用 结果分离出一种新的睾丸特异性基因 显然不符合任何已知的男性生殖细胞- 影响位于 17 号染色体上的基因座。该基因，即蛋白质 其产品含有假定的“锌指”图案，也将 在本提案中详细描述。 我们可能会分离和表征小鼠基因 这里有他们的人类同行。 人类男性的某些形式 可以考虑不孕不育。因为他们的突变。 在这个 从某种意义上说，拟议的项目具有直接的临床相关性并且 应该为我们提供基础知识，使我们能够更好地 人类男性不育症的诊断和治疗。