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

摘要本提案描述了将产生大量拟南芥定位标记的实验。每个标记对应于一个包含限制性内切酶AluI或Sau3A多态性信息的映射DNA片段。该合作项目将有以下目标：(1)通过RFLP和基因组减法克隆包含所需多态性的DNA片段，(2)对克隆进行测序并合成扩增整个片段的PCR引物，(3)确定扩增产物中哪些包含所需多态性，(4)使用GenBank信息生成额外的cap（共显性，裂解扩增多态性序列）。(5)利用重组近交系哥伦比亚X兰斯伯格直立人群体的DNA绘制每个cap图谱；(6)开发能够以有效方式绘制许多基因图谱的方法。***这项工作将为模式植物拟南芥（Arabidopsis thaliana）生成大量的作图标记。经鉴定的克隆序列和有用引物的序列将提供给社区，并将努力安排一家商业公司合成一系列引物，作为廉价的“绘图工具”。如果成功，这项工作将使哥伦比亚或兰茨伯格生态型的任何基因的定位更快、更便宜。% % %

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. %%%