FOX 基因搜寻系统：一种功能获得性基因搜寻的替代技术。

The FOX hunting system: an alternative gain-of-function gene hunting technique.

作者信息

Ichikawa Takanari, Nakazawa Miki, Kawashima Mika, Iizumi Haruko, Kuroda Hirofumi, Kondou Youichi, Tsuhara Yumi, Suzuki Kumiko, Ishikawa Akie, Seki Motoaki, Fujita Miki, Motohashi Reiko, Nagata Noriko, Takagi Tomoko, Shinozaki Kazuo, Matsui Minami

机构信息

Plant Functional Genomics Research Team, Plant Functional Genomics Research Group, Plant Science Center RIKEN Yokohama Institute, 1-7-22 Suehirocho, Tsurumiku, Yokohama, Kanagawa, 230-0045, Japan.

出版信息

Plant J. 2006 Dec;48(6):974-85. doi: 10.1111/j.1365-313X.2006.02924.x.

DOI:10.1111/j.1365-313X.2006.02924.x

PMID:17227551

Abstract

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.

摘要

我们开发了一种新型的功能获得系统，我们将其命名为FOX狩猎系统（全长cDNA过表达基因狩猎系统）。我们使用了标准化的全长cDNA，并通过植物体内转化将每个cDNA导入拟南芥。约10000个独立的拟南芥全长cDNA在拟南芥中CaMV 35S启动子的控制下独立表达。每个转基因拟南芥平均含有2.6个cDNA克隆，并在形态变化、育性和叶色等各类别下进行监测。我们从15547个转化体中发现了1487个可能的形态突变体。当对115个浅绿色T(1)突变体进行分析时，59个株系在超过50%的T(2)后代中表现出突变表型。对两个叶色突变体的表征揭示了这种方法的重要性。我们还记录了来自几个类别的突变体及其相应的全长cDNA。