MOLECULAR BIOLOGY OF CHLOROPLAST RIBOSOMES

叶绿体核糖体的分子生物学

• 批准号: 3274393
• 负责人: DON P BOURQUE
• 金额: $ 11.1万
• 依托单位: UNIVERSITY OF ARIZONA
• 依托单位国家: 美国
• 财政年份: 1979
• 资助国家: 美国
• 起止时间: 1979-09-28 至 1987-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3274393
• 关键词: DNA; chloroplasts; endonuclease; fluoroscopy; gel electrophoresis; genetic mapping; genetic transcription; genetic translation; messenger RNA; nucleic acid denaturation; radiotracer; ribosomes; stoichiometry; tobacco

We propose to study 1) the site of translation of the genes which code for the proteins of chloroplast ribosomes, 2) the intracellular location of these genes, and 3) the physical location of these genes on chloroplast DNA. The site of synthesis of chloroplast ribosomal proteins will be examined by an in vivo mistranslation assay and by translation inhibitor studies. Isolated chloroplast and cytoplasmic fractions will be tested for endogenous synthesis of chloroplast ribosomal proteins in vitro. Messenger RNA fractions from cell cytoplasm and chloroplasts will be examined for ability to program in vitro synthesis of chloroplast ribosomal proteins by translation with 1) E. Coli ribosomes, 2) a reconstituted Nicotiana chloroplast system, 3) wheat germ ribosomes and 4) a reticulocyte lysate system. Isolated fractions with mRNA activity for specific chloroplast ribosomal proteins will allow physical mapping of genes for chloroplast ribosomal proteins. Chloroplast DNA genes will then be physically located by a combination of DNA-RNA hybridization, restriction enzyme mapping procedures, ribosome binding to chloroplast DNA, and coupled transcription-translation studies using individual restriction fragments of chloroplast DNA.

我们建议研究1）编码蛋白质的基因的翻译位点， 叶绿体核糖体的蛋白质，2）细胞内的位置，这些 基因，以及3）这些基因在叶绿体DNA上的物理位置。 的 叶绿体核糖体蛋白质合成位点将通过一种 体内错误翻译测定和翻译抑制剂研究。 分离 将测试叶绿体和细胞质组分的内源性合成， 叶绿体核糖体蛋白。 来自细胞的信使RNA组分 将检查细胞质和叶绿体的体外编程能力 叶绿体核糖体蛋白质的翻译合成：1）E.杆菌 核糖体，2）重组烟草叶绿体系统，3）小麦胚芽 核糖体和4）网织红细胞裂解物系统。 具有mRNA的分离级分 活性的特定叶绿体核糖体蛋白将允许物理映射 叶绿体核糖体蛋白的基因。 叶绿体DNA基因将被 通过DNA-RNA杂交、限制性内切酶 映射程序，核糖体结合叶绿体DNA，并耦合 使用单个限制性片段的转录-翻译研究 叶绿体DNA