Development of Molecular Biology Approaches to Arabidopsis Thaliana

拟南芥分子生物学方法的发展

• 批准号: 8719440
• 负责人: Ronald Davis
• 金额: $ 37.25万
• 依托单位: Stanford University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-05-01 至 1991-10-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=8719440&HistoricalAwards=false
• 关键词: Development; Molecular; Biology; Approaches; Arabidopsis

It is proposed to clone, sequence and map transcriptionally inducible genes from Arabidopsis thaliana. These genes will be the nitrate reductase gene inducible by nitrate; chitinase inducible by ethylene and pathogens, and gibberellic acid inducible genes. The cis acting sites will be mapped by deletion analysis, linker scanning and small fragment insertion into a test promoter. In order to facilitate gene isolation and mapping a complete physical map will be constructed for Arabidopsis. The Arabidopsis chromosomes will be physically separated and isolated using an apparatus applying a contour-clamed homogeneous electric field with a transverse alternating angle of 120o. The NotI fragments from each chromosome will then be separated and correlated with the RFLP genetic map by hybridization with each RFLP probe. Each fragment will also be cloned in a newly developed yeast vector. We will focus attention on the cloning of an Arabidopsis centromere. These will be used in the construction of an artificial chromosome shuttle vector between yeast and Arabidopsis. Arabidopsis thaliana has great advantages over other plants for molecular studies. The results of this work will greatly benefit other investigators using this plant, particularly the integration of the RFLP map with NOTI fragments and the development of an artificial chromosome shuttle vector between yeast and Arabidopsis.

拟对拟南芥转录诱导基因进行克隆、测序和定位。 这些基因将是硝酸盐诱导的硝酸盐还原酶基因；由乙烯和病原体诱导的几丁质酶以及赤霉酸诱导基因。 顺式作用位点将通过缺失分析、接头扫描和小片段插入测试启动子来定位。 为了促进基因分离和作图，将为拟南芥构建完整的物理图谱。 将使用施加横向交错角为 120° 的轮廓均匀电场的装置对拟南芥染色体进行物理分离和分离。 然后，来自每个染色体的 NotI 片段将被分离，并通过与每个 RFLP 探针杂交与 RFLP 遗传图谱相关。 每个片段也将被克隆到新开发的酵母载体中。 我们将重点关注拟南芥着丝粒的克隆。 这些将用于构建酵母和拟南芥之间的人工染色体穿梭载体。 拟南芥在分子研究方面比其他植物具有巨大的优势。 这项工作的结果将极大地惠及使用该植物的其他研究人员，特别是 RFLP 图谱与 NOTI 片段的整合以及酵母和拟南芥之间人工染色体穿梭载体的开发。