The FOX hunting system: an alternative gain-of-function gene hunting technique.
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.
Ichikawa
Takanari
T
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.
Nakazawa
Miki
M
Kawashima
Mika
M
Iizumi
Haruko
H
Kuroda
Hirofumi
H
Kondou
Youichi
Y
Tsuhara
Yumi
Y
Suzuki
Kumiko
K
Ishikawa
Akie
A
Seki
Motoaki
M
Fujita
Miki
M
Motohashi
Reiko
R
Nagata
Noriko
N
Takagi
Tomoko
T
Shinozaki
Kazuo
K
Matsui
Minami
M
D017360:Arabidopsis / Q000235:genetics*; D018076:DNA, Complementary; D018744:DNA, Plant; D015723:Gene Library; D005821:Genetic Techniques; D018745:Genome, Plant; D016296:Mutagenesis; D009857:Oncogenes; D010641:Phenotype; D018515:Plant Leaves / Q000235:genetics; D030821:Plants, Genetically Modified; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D012231:Rhizobium / Q000235:genetics; D014170:Transformation, Genetic
D018076:DNA, Complementary; D018744:DNA, Plant