Molecular Genetic Analysis of the Arabidopsis AB12 Locus

拟南芥 AB12 基因座的分子遗传学分析

• 批准号: 9105241
• 负责人: Ruth Finkelstein
• 金额: $ 24万
• 依托单位: University of California-Santa Barbara
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-08-01 至 1996-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9105241&HistoricalAwards=false
• 关键词: Molecular; Genetic; Analysis; Arabidopsis; AB12

The long term objective of this project is to identify proteins involved in abscisic acid (ABA) action and study their function by physiological, biochemical and molecular genetic analysis. Genes required for ABA action have been identified by isolating and characterizing mutants with altered sensitivity to ABA in Arabidopsis. Dr. Finkelstein's previous studies indicated that three ABI loci (abi1,abi2 and abi3) function in overlapping pathways of ABA response and the loss of function in both pathways was lethal in pollen. She will extend these studies by cloning the relevant loci, either by chromosome walking or genomic subtraction, focussing on ABI2. The specific objectives are: 1) Investigate timing and basis of loss of viability in abi2abi3 pollen, 2)Isolate new radiation induced abi mutants, 3) Isolate overlapping yeast artificial chromosomes spanning the region between the most tightly linked restriction fragment length polymorphism markers flanking the ABI1 locus. Alternatively, if any of the radiation induced mutations are allelic to abi2, she will use genomic subtraction to clone the region deleted in the mutant, 4) Identify the subset of genomic clones showing tightest linkage to ABI2 by RFLP fine mapping these clones with a panel of recombinant families, 5) Identify the ABI2 gene within this subset by complementation of the mutant phenotype, and 6) Isolate and characterize cDNA clones corresponding to the ABI2 gene.

本项目的长期目标是通过生理、生化和分子遗传学分析，鉴定参与脱落酸（ABA）作用的蛋白并研究其功能。通过分离和鉴定拟南芥中对ABA敏感性改变的突变体，确定了ABA作用所需的基因。Finkelstein博士之前的研究表明，三个ABI基因座（abi1，abi2和abi3）在ABA反应的重叠途径中起作用，在花粉中这两个途径的功能丧失是致命的。她将通过染色体行走或基因组减法克隆相关位点来扩展这些研究，重点关注ABI2。具体目标是：1)研究abi2abi3花粉活力丧失的时间和基础；2)分离新的辐射诱导abi突变体；3)分离跨越ABI1位点两侧最紧密连接的限制性片段长度多态性标记之间区域的重叠酵母人工染色体。或者，如果任何辐射诱导的突变与abi2等位，她将使用基因组减法克隆突变中缺失的区域，4)通过RFLP精细定位这些克隆与重组家族面板确定与abi2联系最紧密的基因组克隆子集，5)通过突变表型的互补确定该子集中的abi2基因。6)分离并鉴定ABI2基因对应的cDNA克隆。