Isolation of Arabidopsis Genes by Phenotype Rescue of Seedling Transformants.

我们已经构建了一个转化能力，拟南芥生态型 农杆菌宿主中哥伦比亚菌株GH 50的核基因组文库 菌株AGL 1和粘粒载体pLZO 3，两者都是专门工程化的， 这些目的。 该基因组文库包含约22，000个随机 将克隆在225个96孔微量滴定板上网格化，如果是随机的， 携带大约五个拟南芥基因组等价物。 拟南芥 菌株GH 50携带一个显性乙酰乳酸合酶（als）等位基因 赋予氯磺隆抗性。 我们将发展高 拟南芥全株DNA转化新方法 发芽的幼苗作为基因拯救的工具。 由一个“连续 近似”的方法，我们将进行幼苗转化， 以便能够推断基因组文库中的单个克隆 产生转化体。 我们将利用这种转换- 拟南芥GH50基因组文库构建 农杆菌以拯救赋予抗 磺酰脲类除草剂氯磺隆。 如果成功开发，这项技术将使 探险队分离出植物生长的核心基因， 发展 这种基因拯救技术将使我们能够 高效整合拟南芥物理基因组和遗传 maps.*//

We have constructed a transformation-competent, Arabidopsis ecotype Columbia strain GH50 nuclear genomic library in Agrobacterium host strain AGL1 and cosmid vector pLZO3, both specially engineered for these purposes. This genomic library comprises some 22,000 random clones gridded onto 225 96-well microtiter dishes and, if random, carries some five Arabidopsis genomic equivalents. Arabidopsis strain GH50 carries a dominant acetolactate synthase (als) allele conferring chlorsulfuron resistance. We shall develop high efficiency Arabidopsis whole-plant DNA transformation of newly germinated seedlings as a tool for gene rescue. By a "successive approximation" method, we shall conduct seedling transformation so as to be able to infer individual clones in the genomic library giving rise to transformants. We shall use the transformation- competent Arabidopsis GH50 genomic library constructed in Agrobacterium to rescue the ahs1-1 allele conferring resistance to the sulfonylurea herbicide chlorsulfuron. If successfully developed, this technology will allow the expeditions isolation of genes central to plant growth and development. This gene rescue technology will allow us to efficiency integrate Arabidopsis physical genomic and genetic maps.***//