A single nucleotide polymorphism genotyping method using a phosphate-binding tag nanomolecule

• Project/Area Number: 19590040
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Physical pharmacy
• Research Institution: Hiroshima University
• Principal Investigator: KINOSHITA Eiji Hiroshima University, 大学院・医歯薬学総合研究科, 准教授 (80304418)
• Project Period (FY): 2007 – 2008
• Project Status: Completed (Fiscal Year 2008)
• Budget Amount: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2008: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2007: ¥3,380,000 (Direct Cost: ¥2,600,000、Indirect Cost: ¥780,000)
• Keywords: ゲノム / 遺伝子多型 / 一塩基多型(SNP) / リン酸 / 電気泳動 / フォスタグ(Phos-tag) / エピジェネティックス / メチル化 / 亜鉛 / 遺伝子 / フォスタグ / オーダーメード医療 / 一塩基多型 / SNP

Research Abstract

研究代表者は, リン酸基に選択的な親和性をもつ機能性ナノ分子(フォスタグ, Phos-tag)を用いた新しい一塩基多型(SNP)検出法を開発した。この方法では, Phos-tagを共重合させた電気泳動用ポリアクリルアミドゲルを使用する。この方法により, ミニゲルクラスの電気泳動装置で, ほぼ100%の精度でSNPタイピングができる。本法が, 遺伝子多型の基礎研究分析や臨床診断に貢献できるものと期待している。

Research Products

• [Journal Article] Phosphate-affinity gel electrophoresis using a Phos-tag molecule for phosphoproteome study (2009)
  • Author(s): Eiji Kinoshita, Kinoshita-Kikuta, E., and Koike, T.
  • Journal Title: Curr. Proteomics Proteomics (in press)
  • Peer Reviewed
• [Journal Article] Phos-tag beads as an immunoblotting enhancer for selective detection of phosphoproteins in cell lysates (2009)
  • Author(s): Kinoshita-Kikuta, E., Eiji Kinoshita, and Koike, T.
  • Journal Title: Anal. Biochem (in press)
  • NAID: 120002023644
  • Peer Reviewed
• [Journal Article] A Phos-tag-based fluorescence resonance energy transfer system for the analysis of the dephosphorylation of phosphopeptides (2009)
  • Author(s): Takiyama, K., Eiji Kinoshita, Kinoshita-Kikuta, E., Fujioka, Y., Kubo, Y., and Koike, T.
  • Journal Title: Anal. Biochem (in press)
  • NAID: 120002023643
  • Peer Reviewed
• [Journal Article] Synthetic ciguatoxins selectively activate NAv1. 8-derived chimeric sodium channels expressed in HEK293 cells (2009)
  • Author(s): Yamaoka, K., Inoue, M., Miyazaki, K., Hirama, M., Kondo, C., Eiji Kinoshita, Miyoshi, H., and Seyama
  • Journal Title: J. Biol. Chem. 284 Pages: 7597-7605
  • Peer Reviewed
• [Journal Article] A synthetic approach to develop peptide inhibitors selective for brain-type sodium channels on the basis of pompilidotoxin structure (2009)
  • Author(s): Yokote, S., Setoguchi, R., Shimizu, E., Mishima, N., Kawahara, K., Kuniyasu, A., Shirasaki, T., Takahama, K., Konno, K., Kawai, N., Yamaoka, K., Eiji Kinoshita, and Nakayama, H.
  • Journal Title: Heterocycles 79 Pages: 925-933
  • NAID: 120005255069
  • Peer Reviewed
• [Journal Article] Two-dimensional phosphate-affinity gel electrophoresis for the analysis of phosphoprotein isotypes (2009)
  • Author(s): Eiji Kinoshita, Kinoshita-Kikuta, E., Matsubara, M., Aoki, Y., Ohie, S., Mouri, Y., and Koike, T.
  • Journal Title: Electrophoresis 30 Pages: 550-559
  • Peer Reviewed
• [Journal Article] Two-dimensional phosphate -affinity gel electrophoresis for theanalysis of phosphoprotein isotypes. (2009)
  • Author(s): Kinoshita, Eiji
  • Journal Title: Electrophoresis 30 Pages: 550-559
  • Peer Reviewed
• [Journal Article] A synthetic approach to develop peptide inhibitors selectivefor brain -type sodium channels on the basis of pompilidotoxinstructure. (2009)
  • Author(s): Yokote, Sawana
  • Journal Title: Heterocycles (In press)
  • Peer Reviewed
• [Journal Article] Synthetic ciguatoxins selectively activate NA _v1.8-derived chimeric sodium channels expressed in HEK293 cells (2009)
  • Author(s): Yamaoka, Kaoru
  • Journal Title: J. Biol. Chem. (In press)
  • Peer Reviewed
• [Journal Article] A Phos -tag-based fluorescence resonance energy transfer system for the analysis of the dephosphorylation of phosphopeptides (2009)
  • Author(s): Takiyama, Kei
  • Journal Title: Anal. Biochem. (In press)
  • Peer Reviewed
• [Journal Article] Detection of phosphorylation on large proteins by Western blotting using Phos-tag containing gel. Nat. Protoc (2008)
  • Author(s): Tomida, J., Kitao, H., Eiji Kinoshita, and Takata M.
  • Journal Title: 232(doi : 10. 1038/nprot)
  • Peer Reviewed
• [Journal Article] FANCI phosphorylation functions as a molecular switch to turn on the Fanconi anemia pathway (2008)
  • Author(s): Ishiai, M., Kitao, H., Smogorzewska, A., Tomida, J., Kinomura, A., Uchida, E., Saberi, A., Eiji Kinoshita, Kinoshita-Kikuta, E., Koike, T., Tashiro, S., Elledge, S.J., and Takata, M.
  • Journal Title: Nat. Struct. Mol. Biol 15 Pages: 1138-1146
  • Peer Reviewed
• [Journal Article] Detection of the Gua/Cyt-to-Cyt/Gua mutation in a Gua/Cyt-lined sequence using Zn^<2+>-cyclen polyacrylamide gel electrophoresis (2008)
  • Author(s): Eiji Kinoshita, Kinoshita-Kikuta, E., Yoshimoto, M., and Koike, T.
  • Journal Title: Anal. Biochem 380 Pages: 122-127
  • NAID: 120001092443
  • Peer Reviewed
• [Journal Article] Separation of phosphoprotein isotypes having the same number of phosphate groups using phosphate-affinity SDS-PAGE (2008)
  • Author(s): Eiji Kinoshita, Kinoshita-Kikuta, E., Matsubara, M., Yamada, S., Nakamura, H., Shiro, Y., Aoki, Y., Okita, K., and Koike, T.
  • Journal Title: Proteomics 8 Pages: 2994-3003
  • Peer Reviewed
• [Journal Article] A mobility shift detection method for DNA methylation analysis using phosphate-affinity polyacrylamide gel electrophoresis (2008)
  • Author(s): Kinoshita-Kikuta, E., Eiji Kinoshita, and Koike, T.
  • Journal Title: Anal. Biochem 378 Pages: 102-104
  • NAID: 120001092444
  • Peer Reviewed
• [Journal Article] A mobility shift detection method for DNA methylation analysis using phosphate -affinity polyacrylamide gel electrophoresis (2008)
  • Author(s): Kinoshita-Kikuta, Emiko
  • Journal Title: Anal. Biochem. 378 Pages: 102-104
  • NAID: 120001092444
  • Peer Reviewed
• [Journal Article] Separation of phosphoprotein isotypes having the same number of phosphate groups using phosphate -affinity SDS-PAGE. (2008)
  • Author(s): Kinoshita, Eiji
  • Journal Title: Proteomics 8 Pages: 2994-3003
  • Peer Reviewed
• [Journal Article] Detection of the Gua/Cyt -to-Cyt/Gua mutation in a Gua/Cyt-lined sequence using Zn ^<2+>-cyclen polyacrylamide gelelectrophoresis. (2008)
  • Author(s): Kinoshita, Eiji
  • Journal Title: Anal. Biochem. 380 Pages: 122-127
  • NAID: 120001092443
  • Peer Reviewed
• [Journal Article] FANCI phosphorylation functions as a molecular switch to turn on the Fanconi anemia pathway. (2008)
  • Author(s): Ishiai, Masamichi
  • Journal Title: Nat. Struct. Mol. Biol. 15 Pages: 1138-1146
  • Peer Reviewed
• [Journal Article] Detection of phosphorylation on large proteins by Western blotting using Phos -tag containing gel. (2008)
  • Author(s): Tomida, Junya
  • Journal Title: Nat. Protoc.
  • Peer Reviewed
• [Journal Article] A mobility shift detection method for DNA methylation analysis using Phosphate affinity polyacrylamide gel electroophoresis (2008)
  • Author(s): Kinoshita-Kikuta, E.
  • Journal Title: Anal. Biochem. (印刷中)
  • Peer Reviewed
• [Journal Article] Novel type of phosphate-affinity electrophoresis for protein kinase profiling (2007)
  • Author(s): Eiji Kinoshita, Kinoshita-Kikuta, E., Aoki Y., and Koike, T.
  • Journal Title: Seibutsu Seibutsu-butsuri butsuri-kagaku 51 Pages: 199-206
  • NAID: 10019830592
  • Peer Reviewed
• [Journal Article] Identification on membrane and characterization of phosphoproteins using an alkoxide-bridged dinuclear metal complex as a phosphate binding tag molecule (2007)
  • Author(s): Nakanishi, T., Ando, E., Furuta, M., Eiji Kinoshita, Kinoshita-Kikuta, E., Koike, T., Tsunasawa, S., and Nishimura, O.
  • Journal Title: J. Biomol. Tech. 18 Pages: 278-286
  • Peer Reviewed
• [Journal Article] A single nucleotide polymorphism genotyping method using phosphate-affinity polyacrylamide gel electrophoresis (2007)
  • Author(s): Eiji Kinoshita, Kinoshita-Kikuta, E., and Koike, T.
  • Journal Title: Anal. Biochem 361 Pages: 294-298
  • NAID: 120000878055
  • Peer Reviewed
• [Journal Article] リン酸基アフィニティー電気泳動法を利用した遺伝子診断法 (2007)
  • Author(s): 木下英司, 木下恵美子, 小池透
  • Journal Title: 実験医学 25 Pages: 3033-3039
• [Journal Article] リン酸基アフィニティー電気泳動法を利用した細胞内タンパク質リン酸化反応の解析法 (2007)
  • Author(s): 木下英司, 木下恵美子, 小池透
  • Journal Title: 実験医学 25 Pages: 1229-1235
• [Journal Article] リン酸基親和性クロマトグラフィーを用いた細胞内のリン酸化タンパク質の網羅的精製 (2007)
  • Author(s): 木下英司, 木下恵美子, 小池透
  • Journal Title: バイオテクノロジージャーナル 7 Pages: 217-220
• [Journal Article] リン酸基アフィニティー電気泳動法を利用した細胞内タンパク質リン酸化反応の解析法 (2007)
  • Author(s): 木下英司
  • Journal Title: 実験医学 25 Pages: 1229-1235
• [Journal Article] キナーゼプロファイリングのための新しいリン酸アフィニティー電気泳動法 (2007)
  • Author(s): 木下英司
  • Journal Title: 生物物理化学 51 Pages: 199-206
  • NAID: 10019830592
  • Peer Reviewed
• [Journal Article] 金属イオンの特性を利用したリン酸化タンパク質の特異的認識と検出 (2007)
  • Author(s): 木下英司
  • Journal Title: 薬学雑誌 127 Pages: 1897-1913
  • NAID: 110006546566
  • Peer Reviewed
• [Journal Article] リン酸基アフィニティー電気泳動法を利用した遺伝子診断法 (2007)
  • Author(s): 木下英司
  • Journal Title: 実験医学 25 Pages: 3033-3039
• [Journal Article] Identification on membrane and characterization of phosphoproteins using an alkoxide-bridged dinuclear metal complex as a phosphate binding tag molecule (2007)
  • Author(s): Nakanishi, T.
  • Journal Title: J. Biomol. Tech. 18 Pages: 278-286
  • Peer Reviewed
• [Presentation] リン酸基結合ナノ分子を利用した高感度・高効率なエピジェネティクス変異検出技法の創成 (2009)
  • Author(s): 木下英司, 木下恵美子, 中島弘美, 氏原弘洋, 小池透
  • Organizer: 日本薬学会第129年会
  • Place of Presentation: 国立京都国際会館
  • Year and Date: 2009-03-26
• [Presentation] リン酸基親和性電気泳動法を利用した同じリン酸化量を有するリン酸化蛋白質異性体の分離検出 (2008)
  • Author(s): 木下英司, 木下恵美子, 松原 守, 山田斉爾, 中村寛夫, 城 宜嗣, 青木悠里, 沖田鋼季, 小池透
  • Organizer: BMB2008
  • Place of Presentation: 神戸国際展示場
  • Year and Date: 2008-12-12
• [Presentation] リン酸基親和性電気泳動法を利用した新しい遺伝子診断技術 (2008)
  • Author(s): 木下英司, 木下恵美子, 小池透
  • Organizer: 日本電気泳動学会総会
  • Place of Presentation: 麻布大学百周年記念ホール
  • Year and Date: 2008-11-16
• [Presentation] リン酸基親和性電気泳動法を利用した新しい遺伝子診断技術 (2008)
  • Author(s): 木下英司
  • Organizer: 第59回日本電気泳動学会総会
  • Place of Presentation: 麻布大学
  • Year and Date: 2008-11-16
• [Presentation] フォスタグ結合型新規リン酸基アフィニティービーズを利用した細胞抽出液からのリン酸化蛋白質の分離と濃縮 (2008)
  • Author(s): 木下英司, 木下恵美子, 山田篤志, 井上知香, 小池透
  • Organizer: 日本ヒトプロテオーム機構第6回大会
  • Place of Presentation: ホテル阪急エキスポパーク
  • Year and Date: 2008-07-29
• [Presentation] Phosphate Affinity Gel Electrophoresis Using a Phos-tag Molecule for Phosphoproteome Study in Cancer Cells (2008)
  • Author(s): Eiji Kinoshita
  • Organizer: BIT's Annual World Cancer Congress-2008
  • Place of Presentation: Everbright Convention & Exhibition Center, 上海市, 中華人民共和国
  • Year and Date: 2008-06-12
• [Presentation] ウェスタンブロッティング膜に転写されたリン酸化蛋白質の検出とダイレクト膜上質量分析 (2008)
  • Author(s): 木下英司, 木下恵美子, 中西豪, 安藤英治, 古田大, 綱澤進, 西村紀, 小池透
  • Organizer: 日本薬学会第128年会
  • Place of Presentation: パシフィコ横浜
  • Year and Date: 2008-03-28
• [Presentation] リン酸基アフィニティー電気泳動法を応用した新しい遺伝子診断法の開発 (2008)
  • Author(s): 中島弘美
  • Organizer: 日本薬学会
  • Place of Presentation: パシフィコ横浜
  • Year and Date: 2008-03-28
• [Presentation] フォスタグを用いたリン酸化蛋白質の解析~リン酸基認識素子としての二核錯体とその応用~ (2008)
  • Author(s): 木下英司
  • Organizer: 大阪大学蛋白質研究所セミナー「蛋白質翻訳後修飾」
  • Place of Presentation: ホテル阪急エキスポパーク
  • Year and Date: 2008-01-10
• [Presentation] リン酸化蛋白質解析のための新しい二次元電気泳動法 (2007)
  • Author(s): 木下英司, 木下恵美子, 青木悠里, 大家志織, 毛利由佳, 松原守, 小池透
  • Organizer: BMB2007
  • Place of Presentation: パシフィコ横浜
  • Year and Date: 2007-12-12
• [Presentation] Tohru Koike Phosphate affinity gel electrophoresis using a phosphate-binding tag molecule, Phos-tag, for protein kinase and phosphatse profiling (2007)
  • Author(s): Eiji Kinoshita, Emiko Kinoshita-Kikuta
  • Organizer: The 4th Takeda Science Foundation Symposium on PharmaSciences
  • Place of Presentation: シェラトン都ホテル東京
  • Year and Date: 2007-12-03
• [Presentation] 遺伝子変異を認識する機能性分子を用いた遺伝子診断技術の開発 (2007)
  • Author(s): 木下英司, 木下恵美子, 由元実希, 小池透
  • Organizer: 第58回日本電気泳動学会総会
  • Place of Presentation: 宇部全日空ホテル
  • Year and Date: 2007-11-09
• [Presentation] リン酸化プロテオーム研究の新戦略 (2007)
  • Author(s): 木下英司
  • Organizer: 日本ヒトプロテオーム機構第5回大会
  • Place of Presentation: 日本科学未来館
  • Year and Date: 2007-07-30
• [Book] Single Nucleotide Polymorphisms
  • Author(s): Emiko Kinoshita-Kikuta, Eiji Kinoshita, and Tohru Koike
  • Publisher: Zn(II)-Cyclen Polyacrylamide Gel Electrophoresis for SNP Detection(2d Edition, Chapter 11, Humana Press(in press))
• [Book] Single Nucleotide Polymorphisms
  • Author(s): Eiji Kinoshita, Emiko Kinoshita-Kikuta, and Tohru Koike
  • Publisher: Phosphate-Affinity Polyacrylamide Gel Electrophoresis for SNP Genotyping(2d Edition, Chapter 11, Humana Press(in press))
• [Remarks]
  • URL: http://home.hiroshima-u.ac.jp/tkoike/index.html
• [Remarks]
  • URL: http://home.hiroshima-u.ac.jp/tkoike/research.html