Production and characterization of a large population of cDNA-overexpressing transgenic rice plants using Gateway-based full-length cDNA expression libraries

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  • Hakata Makoto
    Division of Genome and Biodiversity Research, National Institute of Agrobiological Sciences Present address: Hokuriku Research Center, National Agricultural Research Center
  • Nakamura Hidemitsu
    Division of Genome and Biodiversity Research, National Institute of Agrobiological Sciences Present address: Department of Applied Biological Chemistry, The University of Tokyo
  • Iida-Okada Keiko
    Division of Genome and Biodiversity Research, National Institute of Agrobiological Sciences
  • Miyao Akio
    Division of Genome and Biodiversity Research, National Institute of Agrobiological Sciences
  • Kajikawa Mariko
    Division of Genome and Biodiversity Research, National Institute of Agrobiological Sciences
  • Imai-Toki Naoko
    Division of Genome and Biodiversity Research, National Institute of Agrobiological Sciences
  • Pang Jinhuan
    Division of Genome and Biodiversity Research, National Institute of Agrobiological Sciences Present address: State Key Laboratory of Plant Genomics, Institute of Microbiology, Chinese Academy of Sciences
  • Amano Kou
    Division of Genome and Biodiversity Research, National Institute of Agrobiological Sciences Present address: BioResource Center, RIKEN
  • Horikawa Akihiko
    Division of Genome and Biodiversity Research, National Institute of Agrobiological Sciences
  • Tsuchida-Mayama Tomoko
    Division of Genome and Biodiversity Research, National Institute of Agrobiological Sciences
  • Song Jianyu
    Division of Genome and Biodiversity Research, National Institute of Agrobiological Sciences Institute of the Society for Techno-innovation of Agriculture, Forestry and Fisheries
  • Igarashi Motoko
    Division of Genome and Biodiversity Research, National Institute of Agrobiological Sciences Present address: Graduate School of Biological Sciences, Nara Institute of Science and Technology
  • Kitamoto Hiroko K.
    Division of Genome and Biodiversity Research, National Institute of Agrobiological Sciences Present address: Environmental Biofunction Division, National Institute for Agro-Environmental Sciences
  • Ichikawa Takanari
    Plant Science Center, RIKEN Yokohama Institute Present address: Okinawa Institute of Science and Technology
  • Matsui Minami
    Plant Science Center, RIKEN Yokohama Institute
  • Kikuchi Shoshi
    Division of Genome and Biodiversity Research, National Institute of Agrobiological Sciences
  • Nagamura Yoshiaki
    Division of Genome and Biodiversity Research, National Institute of Agrobiological Sciences
  • Hirochika Hirohiko
    Division of Genome and Biodiversity Research, National Institute of Agrobiological Sciences
  • Ichikawa Hiroaki
    Division of Genome and Biodiversity Research, National Institute of Agrobiological Sciences Division of Plant Sciences, National Institute of Agrobiological Sciences

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We applied the full-length cDNA overexpressor (FOX) gene-hunting system for systematic and genome-wide functional analysis of rice genes. In this study, we constructed a novel binary vector carrying the Gateway site-specific recombination cassette and then constructed rice FOX libraries containing a maximum of 13,823 independent, full-length cDNAs (fl-cDNAs) that correspond to approximately half the total number of rice fl-cDNA clones. By introducing the FOX libraries via Agrobacterium, we generated 2,586 FOX-rice lines exhibiting various visible alterations (e.g., plant height, tillers, leaves, and heading dates). The introduced fl-cDNAs, integrated into individual transgenic rice genomes, were amplified by genomic PCR and identified using sequencing analysis. The fl-cDNAs were PCR-amplified in 2,251 (94.2%) of the 2,389 FOX-rice lines that were examined, identifying 1,920 independent fl-cDNAs in the FOX lines. In addition to the previously generated FOX-rice plants, our new collection of FOX-rice lines produced through the Gateway system should be a useful tool for the efficient identification of gene functions in rice. Moreover, this Gateway-based technology should be applicable to other species in which a collection of fl-cDNA clones is available.<br>

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