Development of series of gateway binary vectors, pGWBs, for realizing efficient construction of fusion genes for plant transformation

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概要

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• We developed a new series of binary vectors useful for Gateway cloning to facilitate transgenic experiments in plant biotechnology. The new system, Gateway Binary Vectors (pGWBs) realized efficient cloning, constitutive expression using the cauliflower mosaic virus (CaMV) 35S promoter and the construction of fusion genes by simple clonase reaction with an entry clone. The reporters employable in this system are GUS, sGFP, LUC, EYFP, and ECFP. The tags available are 6xHis, FLAG, 3xHA, 4xMyc, 10xMyc, GST, T7-epitope, and tandem affinity purification (TAP). In total, 13 kinds of reporter or tag were arranged and were almost applicable to both N- and C-fusions. The pGWBs could be used for many purposes, such as promoter::reporter analysis, observation of subcellular localization by the expression of proteins fused to a reporter or tag, and analysis of protein-protein interaction by copurification and immunodetection experiments. The pGWBs were constructed with modified pBI101 containing a CaMV35S promoter-driven hygromycin phosphotransferase (HPT) gene as the second selection marker. We also constructed pGWBs with the marker HPT driven by the nopaline synthase promoter. By using the pGWB system, the expression of tagged proteins, and the localization of GFP-fused proteins were easily analyzed. Moreover, tissue-specific and inducible gene expression using a promoter was also monitored with pGWBs. It is expected that, the pGWB system will serve as a powerful tool for plasmid construction in plant research.

• 社団法人日本生物工学会の論文
• 2007-07-25

著者

• NAKAGAWA Tsuyoshi

  Department of Molecular and Functional Genomics, Center for Integrated Research in Science, Shimane

• Niwa Yasuo

  Plant Science Center Riken(the Institute Of Physical And Chemical Research)

• Niwa Yasuo

  Lab. Of Plant Cell Technol. Graduate School Of Nutritional And Environmental Sciences Univ. Of Shizu

• MATSUOKA Ken

  Plant Science Center, RIKEN

• Takahashi N

  Plant Functions Laboratory Riken

• Tanaka K

  Institute Of Molecular And Cellular Biosciences The University Of Tokyo

• Tanaka Katsunori

  Department of Applied Bioscience and Biotechnology, Faculty of Life and Environmental Science, Shima

• Tanaka K

  Tottori Univ. Tottori

• Matsuoka Ken

  Plant Science Center Riken(the Institute Of Physical And Chemical Research):laboratory Of Plant Nutr

• Kurose Takayuki

  Department of Molecular and Functional Genomics, Center for Integrated Research in Science, Shimane

• Hino Takeshi

  Department of Molecular and Functional Genomics, Center for Integrated Research in Science, Shimane

• Kawamukai Makoto

  Laboratory of Plant Cell Technology, Graduate School of Nutritional and Environmental Sciences, Univ

• Toyooka Kiminori

  Laboratory of Plant Nutrition, Faculty of Agriculture, Kyushu University

• Jinbo Tetsuro

  Department of Sustainable Resource Science, Faculty of Bioresources, Mie University

• Kimura Tetsuya

  Plant Science Center, RIKEN(The Institute of Physical and Chemical Research)

• Matsuoka Ken

  Plant Science Center Riken

• Nakano Takeshi

  Riken Advanced Sci. Inst.

• Kimura Tetsuya

  Department Of Sustainable Resource Science Graduate School Of Of Bioresources Mie University

• Takahashi Nobutaka

  Frontier Research Program The Institute Of Physical And Chemical Research

• Takahashi Nobutaka

  Frontier Research Program The Institute Of Physical And Chemical Research (rlken)

• Nakagawa Tetsuto

  Department Of Bioscience And Biotechnology Division Of Bioresource And Bioenvironmental Sciences Gra

• Nakagawa Tsuyoshi

  Dep. Of Molecular And Functional Genomics Center For Integrated Res. In Sci. Shimane Univ.

• Tanaka Kiyoshi

  Coll. Environ. Health Azabu Univ.

• NAKANO Takeshi

  The Institute of Physical and Chemical Research (RIKEN)

• Hino Takeshi

  Department Of Molecular And Functional Genomics Center For Integrated Research In Science Shimane Un

• Jinbo Tetsuro

  Department Of Sustainable Resource Science Faculty Of Bioresources Mie University

• Kimura Tetsuya

  Dep. Of Sustainable Resource Sci. Graduate School Of Bioresources Mie Univ.

• Nakagawa T

  Department Of Bioscience And Biotechnology Division Of Bioresource And Bioenvironmental Sciences Gra

• Nakagawa Tsuyoshi

  Department Of Molecular And Functional Genomics Center For Integrated Research In Science Shimane Un

• Morita K

  Department Of Applied Bioscience And Biotechnology Faculty Of Life And Environmental Science Shimane

• Kawamukai M

  Department Of Applied Bioscience And Biotechnology Faculty Of Life And Environmental Science Shimane

• Takahashi Nobuyuki

  Hokkaido University Of Education

• Kawamukai Makoto

  Department Of Life Science And Biotechnology Faculty Of Life And Environmental Science Shimane Unive

• Toyooka Kiminori

  Riken Yokohama Jpn

• Nakano Takeshi

  Plant Functions Lab. The Inst. Of Physical And Chemical Res.

• Nakano Takeshi

  Plant Functions Lab.

• Kikuchi Takao

  Coll. Environ. Health Azabu Univ.

• Tanaka Keiko

  Department Of Food Science And Technology Faculty Of Agriculture Kyoto University

• Nakagawa Tsuyoshi

  Department Of Molecular And Functional Genomics Center For Integrated Research In Science Shimane Un

• Kawamukai Makoto

  Department Of Life Science And Biotechnology Faculty Of Life And Environmental Science Shimane Unive

• Takahashi Nobuyuki

  Laboratory For Molecular Function Of Food Division Of Food Science And Biotechnology Graduate School

• Koyanagi Takashi

  Research Institute For Bioresources And Biotechnology Ishikawa Prefectural University

• Takahashi Noriko

  Department of Molecular and Functional Genomics, Center for Integrated Research in Science, Shimane University

• Tanaka Kyoko

  Graduate School of Life and Environmental Sciences, Kyoto Prefectural University

• NAKAGAWA Tsuyoshi

  Department of Materials Science and Engineering, Graduate School of Engineering, Nagoya Institute of Technology

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