Immunoproteomic and Two-Dimensional Difference Gel Electrophoresis Analysis of Arabidopsis Dehydration Response Element-Binding Protein 1A (DREB1A)-Transgenic Potato
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概要
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To produce crops that are more tolerant to stresses such as heat, cold, and salt, transgenic plants have been produced those express stress-associated proteins. In this study, we used immunoproteomic and two-dimensional difference gel electrophoresis (2D-DIGE) methods to investigate the allergenicity of transgenic potatoes expressing Arabidopsis DREB1A (dehydration responsive element-binding protein 1A), driven by the rd29A promoter or the 35S promoter. Immunoproteomic analysis using sera from potato-allergic patients revealed several immunoglobulin E (IgE)-binding protein spots. The patterns of protein binding were almost the same between transgenic and non-transgenic potatoes. The IgE-binding proteins in potato were identified as patatin precursors, a segment of serine protease inhibitor 2, and proteinase inhibitor II by matrix assisted laser desorption/ionization-time of flight (MALDI-TOF) MS/MS. 2D-DIGE analysis revealed several differences in protein expression between non-transgenic potato and transgenic potato; those showing increased expression in transgenic potatoes were identified as precursors of patatin, a major potato allergen, and those showing decreased expression in transgenic potatoes were identified as lipoxygenase and glycogen (starch) synthase. These results suggested that transgenic potatoes may express slightly higher levels of allergens, but their IgE-binding patterns were almost the same as those of control potatoes. Further research on changes in protein expressions in response to environmental factors is required to confirm whether the differences observed in this study are due to gene transfection, rather than environmental factors.
著者
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KIKUCHI Akira
Gene Research Center and Graduate School of Life and Environmental Sciences, University of Tsukuba
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SHIMAZAKI Takayoshi
Gene Research Center and Graduate School of Life and Environmental Sciences, University of Tsukuba
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WATANABE Kazuo
Gene Research Center and Graduate School of Life and Environmental Sciences, University of Tsukuba
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Teshima Reiko
Division of Biochemistry and Immunochemistry, National Institute of Health Sciences
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Nakamura Ryosuke
Division of Biochemistry and Immunochemistry, National Institute of Health Sciences
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Nakamura Rika
Division of Biochemistry and Immunochemistry, National Institute of Health Sciences
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SATOH Rie
Division of Novel Foods and Immunochemistry, National Institute of Health Sciences (NIHS)
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Nakamura R
Division Of Novel Foods And Immunochemistry National Institute Of Health Sciences (nihs)
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Kikuchi Akira
Gene Res. Center Graduate School Of Life And Environmental Sciences Univ. Of Tsukuba
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Teshima Reiko
Division Of Biochemistry And Immunochemistry National Institute Of Health Sciences
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Yamaguchi-Shinozaki Kazuko
Biological Resources Division, Japan International Research Center for Agricultural Sciences(JIRCAS)
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KASUGA Mie
Biological Resources Division, Japan International Research Center for Agricultural Sciences (JIRCAS
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Kasuga Mie
Biological Resources Div. Japan International Res. Center For Agricultural Sciences (jircas)
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Satoh Rie
National Inst. Of Health Sciences
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Yamaguchi-shinozaki Kazuko
Biological Resources Division Japan International Research Center For Agicultural Sciences (jircas)
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Watanabe Kazuo
Gene Res. Center Graduate School Of Life And Environmental Sciences Univ. Of Tsukuba
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Yamaguchi-Shinozaki Kazuko
Biological Resources Division, Japan International Research Center for Agricultural Sciences (JIRCAS)
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Kasuga Mie
Biological Resources Division, Japan International Research Center for Agricultural Sciences (JIRCAS)
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Satoh Rie
Division of Novel Foods and Immunochemistry, National Institute of Health Sciences
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Shimazaki Takayoshi
Gene Research Center, Graduate School of Life and Environmental Sciences, University of Tsukuba
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Nakamura Rika
Division of Novel Foods and Immunochemistry, National Institute of Health Sciences
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