The FOX hunting system: an alternative gain-of-function gene hunting technique
The FOX hunting system: an alternative gain-of-function gene hunting technique
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DOI：
10.1111/j.1365-313x.2006.02924.x
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发表时间：
2006-12-01
期刊：
影响因子：
7.2
通讯作者：
Matsui, Minami
中科院分区：
文献类型：
--
作者：
Ichikawa, Takanari;Nakazawa, Miki;Matsui, Minami
我们开发了一种新的功能获得系统，我们命名为FOX狩猎系统（全长cDNA过度表达基因狩猎系统）。我们使用标准化的全长cDNA，并通过植物转化将每个cDNA引入拟南芥。在CaMV 35 S启动子的调控下，约有10000个拟南芥全长cDNA在拟南芥中得到了独立表达。每个转基因拟南芥平均含有2.6个cDNA克隆，并在各种类别下进行监测，如形态变化，生育力和叶色。从15547个转化子中筛选出1487个可能的形态突变体。当分析115个淡绿色T-1突变体时，59个株系代表超过50%的T-2后代的突变表型。两个叶色突变体的表征揭示了这种方法的意义。我们还记录了几类突变体及其相应的全长cDNA。
We have developed a novel gain-of-function system that we have named the FOX hunting system (Full-length cDNA Over-eXpressing gene hunting system). We used normalized full-length cDNA and introduced each cDNA into Arabidopsis by in planta transformation. About 10 000 independent full-length Arabidopsis cDNAs were expressed independently under the CaMV 35S promoter in Arabidopsis. Each transgenic Arabidopsis contained on average 2.6 cDNA clones and was monitored under various categories such as morphological changes, fertility and leaf color. We found 1487 possible morphological mutants from 15 547 transformants. When 115 pale green T-1 mutants were analyzed, 59 lines represented the mutant phenotypes in more than 50% of the T-2 progeny. Characterization of two leaf color mutants revealed the significance of this approach. We also document mutants from several categories and their corresponding full-length cDNAs.