MAIZE RIBOSOMAL PROTEIN GENE STRUCTURE AND EXPRESSION

玉米核糖体蛋白基因结构和表达

• 批准号: 3466439
• 负责人: JAMES S GANTT
• 金额: $ 0.16万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-09-01 至 1994-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3466439
• 关键词: DNA footprinting; chloroplasts; computer data analysis; corn; developmental genetics; gene expression; genetic library; genetic transcription; messenger RNA; mutant; nucleic acid hybridization; nucleic acid sequence; plant genetics; plant growth /development; plant proteins; precursor mRNA; protein biosynthesis; ribosomal RNA; ribosomes

The long-term objectives of the proposed research are to understand how gene structure relates to gene expression; how the cell regulates the expression of large sets of genes to produce functional plastid (70S) and cytosolic (80S) ribosomes; and how the expression of plastid- and nuclear-encoded 70S ribosomal protein genes is coordinated in the separate genomes. A large increase in the cellular content of 70S and 80S ribosomes occurs during maize leaf cell development. The specific aims are to thoroughly characterize the ribosome content throughout maize leaves; determine the content of 80S and plastid-encoded and nuclear- encoded 70S ribosomal protein mRNAs in leaves using ribosomal protein cDNAs as hybridization probes; and to examine the rates of ribosomal protein synthesis in the leaves. Additionally, the influence of 70S ribosome content on the expression nuclear- encoded 70S ribosomal protein gene expression will be studied in mutants of maize which pre cold-sensitive for the accumlation of the 70S ribosomes. These experiments are designed to determine the mechanisms leading to the balanced accumulation of 70S ribosomal proteins. Complementary to the above studies will be the investigation of 80S and nuclear-encoded 70S ribosomal protein gene structure. cDNAs for these genes be isolated using heterologous 70S and 80S ribosomal protein cDNAs. The maize cDNAs will then be used in the above experiments and to select the genomic clones from a maize library. Several 80S and nuclear-encoded 70S ribosomal protein genes will be sequenced. Transcription start and stop sites will be identified by S1 nuclease mapping and primer extension analysis. Computer-assisted sequence analysis of these genes may reveal DNA sequences that could be involved in the coordinate regulation of the ribosomal protein genes. Protein binding activity of the genes will be examined by DNA footprint analysis. Additionally, the possibility that some ribosomal proteins bind to pre-mRNAs or mature mRNAs will be tested by protein-RNA binding assays.

拟议研究的长期目标是 了解基因结构与基因表达的关系；如何 细胞调节大量基因的表达以产生 功能质体 (70S) 和胞质 (80S) 核糖体；以及如何 质体和核编码的 70S 核糖体蛋白的表达 基因在不同的基因组中协调。 大幅增加 70S 和 80S 核糖体的细胞含量发生在玉米过程中 叶细胞发育。 具体目标是彻底 表征整个玉米叶中的核糖体含量； 确定 80S 和质体编码和核的含量 使用核糖体编码叶子中的 70S 核糖体蛋白 mRNA 蛋白质 cDNA 作为杂交探针；并检查费率 叶子中核糖体蛋白质的合成。 此外， 70S核糖体含量对核表达的影响 编码70S核糖体蛋白的基因表达将在 玉米突变体对冷敏感，可积累 70S核糖体。 这些实验旨在确定 70S均衡积累的机制 核糖体蛋白。 对上述研究的补充将是对 80S和核编码的70S核糖体蛋白基因结构。 使用异源 70S 和 80S 分离这些基因的 cDNA 核糖体蛋白 cDNA。 玉米 cDNA 将被用于 进行上述实验并从基因组克隆中选择 玉米图书馆。 几个80S和核编码的70S核糖体 蛋白质基因将被测序。 转录开始和停止 位点将通过 S1 核酸酶作图和引物来识别 延伸分析。 计算机辅助序列分析这些 基因可能揭示可能参与的DNA序列 核糖体蛋白基因的协调调节。 蛋白质 基因的结合活性将通过 DNA 足迹进行检查 分析。 此外，某些核糖体的可能性 与前体 mRNA 或成熟 mRNA 结合的蛋白质将通过以下方式进行测试 蛋白质-RNA 结合测定。