Collaborative Research: A Rapid and Inexpensive Whole-Genome Mapping Method for Arabidopsis Using CAPS Co-Dominant Ecotype-Specific PCR-Based Markers

合作研究：使用 CAPS 共显性生态型特异性 PCR 标记的拟南芥全基因组作图方法

• 批准号: 9406240
• 负责人: Ronald Davis
• 金额: $ 16万
• 依托单位: Stanford University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-08-01 至 1997-01-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9406240&HistoricalAwards=false
• 关键词: Collaborative; Research; Rapid; Inexpensive; Whole

9405961 Abstract This proposal describes experiments that will generate a large set of mapping markers for Arabidopsis thaliana. Each marker will correspond to a mapped DNA fragment that contains an informative polymorphism for the restriction endonucleases AluI or Sau3A. This collaborative project will have the following goals: (1) to clone DNA fragments that contain the desired polymorphisms by RFLP and genomic subtraction, (2) to sequence the clones and synthesize PCR primers that will amplify the entire fragment, (3) to determine which of the amplified products contain the desired polymorphisms, (4) to use GenBank information to generate additional CAPS (co- dominant, cleaved amplified polymorphic sequences), (5) to map each of the CAPS using DNA from a recombinant inbred Columbia X Landsberg erecta population, and (6) to develop methods that will allow many genes to be mapped in an efficient manner. *** This work will generate a large set of mapping markers for the model plant, Arabidopsis thaliana. The sequences of the clones and of useful primers, as they are identified, will be provided to the community and efforts will be made to arrange for a commercial firm to synthesize sets of primes as inexpensive "mapping kits". If successful, this work should make mapping of any gene from either the Columbia or the Landsberg ecotype much, much faster and much less expensive. %%%

9405961摘要本提案描述了将为拟南芥产生大量作图标记的实验。每个标记将对应于一个映射的DNA片段，该片段包含限制内切酶Alu1或Sau3A的信息性多态。该合作项目将有以下目标：(1)通过RFLP和基因组消减来克隆包含所需多态的DNA片段，(2)对克隆进行测序并合成将扩增整个片段的PCR引物，(3)确定哪些扩增产物包含所需的多态，(4)利用GenBank的信息来生成额外的CAP(共显性的、切割的扩增的多态序列)，(5)使用来自重组近交系哥伦比亚X Landsberg erecta群体的DNA绘制每个CAP的图谱，以及(6)开发能够以高效的方式对许多基因进行图谱的方法。*这项工作将为模式植物拟南芥产生大量的作图标记。克隆和有用的引物的序列一旦确定，将被提供给社区，并将努力安排一家商业公司合成几套引物，作为廉价的“测绘工具包”。如果成功，这项工作将使哥伦比亚生态型或兰茨堡生态型的任何基因的作图变得非常非常快，而且成本也低得多。%%%