Development of Gateway Binary Vectors, R4L1pGWBs, for Promoter Analysis in Higher Plants

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  • NAKAMURA Shinya
    Department of Molecular and Functional Genomics, Center for Integrated Research in Science, Shimane University
  • NAKAO Akihide
    Department of Molecular and Functional Genomics, Center for Integrated Research in Science, Shimane University
  • KAWAMUKAI Makoto
    Department of Applied Bioscience and Biotechnology, Faculty of Life and Environmental Science, Shimane University
  • KIMURA Tetsuya
    Department of Sustainable Resource Science, Graduate School of Bioresources, Mie University
  • ISHIGURO Sumie
    Department of Biological Mechanisms and Functions, Graduate School of Bioagricultural Sciences, Nagoya University
  • NAKAGAWA Tsuyoshi
    Department of Molecular and Functional Genomics, Center for Integrated Research in Science, Shimane University

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Abstract

We developed a new series of Gateway binary vectors for plant transformation, R4L1pGWBs, which allow easy construction of promoter:reporter clones. R4L1pGWBs contain a recombination attR4-attL1-reporter cassette, and thus an attL4-promoter-attR1 entry clone was efficiently incorporated by the Gateway LR reaction, resulting in the generation of an attB4-promoter-attB1-reporter construct. The reporters employed in R4L1pGWBs were β-glucuronidase (GUS), luciferase (LUC), enhanced yellow fluorescent protein (EYFP), enhanced cyan fluorescent protein (ECFP), G3 green fluorescent protein (G3GFP), G3GFP-GUS, and tag red fluorescent protein (TagRFP).

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