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Study on exploration of new crop-specific and multi-copied DNA markers using bioinformatic approach

Research Project

Project/Area Number 20K05901
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Review Section Basic Section 38050:Food sciences-related
Research InstitutionNational Institute of Health Sciences

Principal Investigator

Soga KEISUKE  国立医薬品食品衛生研究所, 生化学部, 主任研究官 (50746336)

Co-Investigator(Kenkyū-buntansha) 吉田 光範  国立感染症研究所, ハンセン病研究センター 感染制御部, 主任研究官 (70772630)
Project Period (FY) 2020-04-01 – 2024-03-31
Project Status Completed (Fiscal Year 2023)
Budget Amount *help
¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000)
Fiscal Year 2022: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2021: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2020: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Keywordsバイオインフォマティクス / 遺伝子検査 / リアルタイムPCR / コメ / とうもろこし / マルチコピー / DNAマーカー / 食品分析 / マルチコピーDNA / 遺伝子マーカー / トウモロコシ / 加工食品 / セントロメア / リピート配列 / リボソームDNA / ITS / 検出法 / ゲノムワイド
Outline of Research at the Start

加工食品には様々な原材料が使われているが、加工工程が複雑なほどタンパク質やDNAが分解・断片化され、その検出は困難である。そこで、本研究では、原材料に特異的でマルチコピーのDNA配列を探索し、それを標的とした加工食品中の作物単独または一斉検出法を開発することを目的とする。既知のマルチコピーDNA(リボソームRNAまたはトランスポゾン領域など)の作物種間マルチアライメントを行う手法、ゲノムワイドに新規マルチコピー配列を探索する手法をそれぞれ検討し、候補配列を列挙する。そして、リアルタイムPCRまたは次世代シーケンサー等を用いたメタバーコーディング法を開発し、加工食品への適用性を検証する。

Outline of Final Research Achievements

Genetic testing methods such as PCR targeting species-specific and multi-copy DNA markers are useful for detecting adulteration, contamination in processed foods and crops in vomit.
In this study, we investigated the maize-specific and multi copied markers targeting ribosomal RNA. We also investigated the rice-specific and multi copied markers using bioinformatics approaches, and a search pipeline was developed. Highly sensitive real-time PCRs were successfully developed by targeting the respective candidate marker regions. On the other hand, non-specific and trace amplification could not be completely eliminated in certain crops, and further studies are required when applying them to tests.

Academic Significance and Societal Importance of the Research Achievements

本研究で開発された高感度リアルタイムPCR法を用いることで、加工食品中のとうもろこしおよびイネの混入の確認が可能と考えられる。この場合は、食品偽装食品中のアレルゲン性の確認等の検査への要用が期待できる。また、高感度かつ特異的マーカー探索パイプラインは、今回検討した作物以外の作物への応用も期待でき、食品遺伝子検査の高感度化に貢献できると考えられる。

Report

(5 results)
  • 2023 Annual Research Report   Final Research Report ( PDF )
  • 2022 Research-status Report
  • 2021 Research-status Report
  • 2020 Research-status Report
  • Research Products

    (24 results)

All 2023 2022 2021 2020

All Journal Article (11 results) (of which Peer Reviewed: 9 results,  Open Access: 5 results) Presentation (13 results) (of which Int'l Joint Research: 2 results)

  • [Journal Article] Investigation of Genetically Modified Maize Imported into Japan in 2021/2022 and the Applicability of Japanese Official Methods2023

    • Author(s)
      Soga K, Taguchi C, Sugino M, Egi T, Narushima J, Yoshiba S, Takabatake R, Kondo K, Shibata N
    • Journal Title

      Food Hygiene and Safety Science (Shokuhin Eiseigaku Zasshi)

      Volume: 64 Issue: 6 Pages: 218-225

    • DOI

      10.3358/shokueishi.64.218

    • ISSN
      0015-6426, 1882-1006
    • Year and Date
      2023-12-25
    • Related Report
      2023 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Simple and quick detection of extended-spectrum <i>β</i>-lactamase and carbapenemase-encoding genes using isothermal nucleic acid amplification techniques2023

    • Author(s)
      NAKAYAMA TATSUYA、SOGA KEISUKE
    • Journal Title

      Journal of Microorganism Control

      Volume: 28 Issue: 4 Pages: 145-152

    • DOI

      10.4265/jmc.28.4_145

    • ISSN
      2758-6383, 2758-6391
    • Related Report
      2023 Annual Research Report
  • [Journal Article] Providing appropriate information to consumers boosts the acceptability of genome-edited foods in Japan2023

    • Author(s)
      Taguchi Chie、Shibata Norihito、Soga Keisuke、Yoshiba Satoko、Narushima Jumpei、Sugino Miyu、Kondo Kazunari
    • Journal Title

      GM Crops &amp; Food

      Volume: 14 Issue: 1 Pages: 1-14

    • DOI

      10.1080/21645698.2023.2239539

    • Related Report
      2023 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Evaluation of Conversion Factor for Rapid Quantification of Authorized Genetically Modified Maize and Soybean in Japan2022

    • Author(s)
      Shibata Norihito、Soga Keisuke、Sugino Miyu、Narushima Jumpei、Yoshiba Satoko、Egi Tomohiro、Takabatake Reona、Kondo Kazunari
    • Journal Title

      BPB Reports

      Volume: 5 Issue: 5 Pages: 115-120

    • DOI

      10.1248/bpbreports.5.5_115

    • ISSN
      2434-432X
    • Related Report
      2022 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Unbiased prediction of off‐target sites in genome‐edited rice using <scp>SITE‐Seq</scp> analysis on a web‐based platform2022

    • Author(s)
      Narushima Jumpei、Kimata Shinya、Shiwa Yuh、Gondo Takahiro、Akimoto Satoshi、Soga Keisuke、Yoshiba Satoko、Nakamura Kosuke、Shibata Norihito、Kondo Kazunari
    • Journal Title

      Genes to Cells

      Volume: 27 Issue: 12 Pages: 706-718

    • DOI

      10.1111/gtc.12985

    • Related Report
      2022 Research-status Report
    • Peer Reviewed
  • [Journal Article] Development and Validation of a New Robust Detection Method for Low-Content DNA Using ΔΔCq-Based Real-Time PCR with Optimized Standard Plasmids as a Control Sample2022

    • Author(s)
      Soga Keisuke、Nakamura Kosuke、Egi Tomohiro、Narushima Jumpei、Yoshiba Satoko、Kishine Masahiro、Mano Junichi、Kitta Kazumi、Takabatake Reona、Shibata Norihito、Kondo Kazunari
    • Journal Title

      Analytical Chemistry

      Volume: 94 Issue: 41 Pages: 14475-14483

    • DOI

      10.1021/acs.analchem.2c03680

    • Related Report
      2022 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Development and Interlaboratory Validation of a Novel Reproducible Qualitative Method for GM Soybeans Using Comparative Cq-Based Analysis for the Revised Non-GMO Labeling System in Japan2022

    • Author(s)
      Takabatake Reona、Egi Tomohiro、Soga Keisuke、Narushima Jumpei、Yoshiba Satoko、Shibata Norihito、Nakamura Kosuke、Kondo Kazunari、Kishine Masahiro、Mano Junichi、Kitta Kazumi
    • Journal Title

      Analytical Chemistry

      Volume: 94 Issue: 39 Pages: 13447-13454

    • DOI

      10.1021/acs.analchem.2c02447

    • Related Report
      2022 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Strategy for detecting off-target sites in genome-edited rice2021

    • Author(s)
      Jumpei Narushima, Shinya Kimata, Yuh Shiwa, Takahiro Gondo, Satoru Akimoto, Keisuke Soga, Satoko Yoshiba, Kosuke Nakamura, Norihito Shibata, Kazunari Kondo
    • Journal Title

      bioRxiv

      Volume: 5/29 Pages: 1-26

    • DOI

      10.1101/2021.05.28.446070

    • Related Report
      2021 Research-status Report
    • Open Access
  • [Journal Article] Development and Testing of an Individual Kernel Detection System for Genetically Modified Soybean Events in Non-identity-preserved Soybean Samples2020

    • Author(s)
      Soga Keisuke、Kimata Shinya、Narushima Jumpei、Sato Sakiko、Sato Emi、Mano Junichi、Takabatake Reona、Kitta Kazumi、Kawakami Hiroshi、Akiyama Hiroshi、Kondo Kazunari、Nakamura Kosuke
    • Journal Title

      Biological and Pharmaceutical Bulletin

      Volume: 43 Issue: 8 Pages: 1259-1266

    • DOI

      10.1248/bpb.b20-00382

    • NAID

      130007883796

    • ISSN
      0918-6158, 1347-5215
    • Year and Date
      2020-08-01
    • Related Report
      2020 Research-status Report
    • Peer Reviewed
  • [Journal Article] Evaluation of Conversion Factors for Genetically Modified Maize Quantification2020

    • Author(s)
      Takabatake Reona、Onishi Mari、Mano Junichi、Kishine Masahiro、Soga Keisuke、Nakamura Kosuke、Kondo Kazunari、Kitta Kazumi
    • Journal Title

      Food Hygiene and Safety Science (Shokuhin Eiseigaku Zasshi)

      Volume: 61 Issue: 6 Pages: 235-238

    • DOI

      10.3358/shokueishi.61.235

    • NAID

      130007960250

    • Related Report
      2020 Research-status Report
    • Peer Reviewed
  • [Journal Article] Development of a Novel Detection Method Targeting an Ultrashort 25 bp Sequence Found in Agrobacterium-Mediated Transformed GM Plants2020

    • Author(s)
      Takabatake Reona、Onishi Mari、Minegishi Yasutaka、Futo Satoshi、Soga Keisuke、Nakamura Kosuke、Kondo Kazunari、Mano Junichi、Kitta Kazumi
    • Journal Title

      Journal of Agricultural and Food Chemistry

      Volume: 68 Issue: 51 Pages: 15327-15334

    • DOI

      10.1021/acs.jafc.0c03864

    • Related Report
      2020 Research-status Report
    • Peer Reviewed
  • [Presentation] For the development of a PCR primer design pipeline for detection of contamination in foods.2023

    • Author(s)
      Yoritaka Harazono, Keisuke Soga, Masahiro Kasahara.
    • Organizer
      Genome Informatics
    • Related Report
      2023 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Development and validation of a new robust qualitative method for GM maize using comparative Cq-based real-time PCR for the revised non-GM labeling system in Japan.2023

    • Author(s)
      Soga, K., Egi, T., Narushima, J., Yoshiba, S., Kishine, M., Mano, J., Kitta, K., Takabatake, R., Adachi, R., Nakamura, K., Kondo, K., Shibata, N.
    • Organizer
      AOAC 137th Annual Meeting and Exposition
    • Related Report
      2023 Annual Research Report
    • Int'l Joint Research
  • [Presentation] バイオインフォマティクスツールを活用した種特異的遺伝子配列探索法の開発2023

    • Author(s)
      杉野御祐、成島純平、曽我慶介、吉場聡子、田口千恵、安達玲子、柴田識人
    • Organizer
      第46回日本分子生物学会
    • Related Report
      2023 Annual Research Report
  • [Presentation] 遺伝子検査法における種特異的遺伝子探索のためのバイオインフォマティクスの活用2023

    • Author(s)
      杉野御祐、成島純平、曽我慶介、吉場聡子、田口千恵、安達玲子、柴田識人
    • Organizer
      第9回次世代を担う若手のためのレギュラトリーサイエンスフォーラム
    • Related Report
      2023 Annual Research Report
  • [Presentation] 遺伝子組換えとうもろこし系統の国内流通実態調査(2021年度産)2023

    • Author(s)
      曽我慶介、田口千恵、杉野御祐、江木智宏、成島純平、吉場聡子、高畠令王奈、安達玲子、近藤一成、柴田識人
    • Organizer
      日本食品衛生学会第119回学術講演会
    • Related Report
      2023 Annual Research Report
  • [Presentation] Cas9 targeted long-read sequencingによる食品中の外来性遺伝子配列の同定2023

    • Author(s)
      成島純平、吉場聡子、細川葵、曽我慶介、杉野御佑、田口千恵、安達玲子、近藤一成、柴田識人
    • Organizer
      第46回日本分子生物学会
    • Related Report
      2023 Annual Research Report
  • [Presentation] ダイズ穀粒における遺伝子組換え農産物混入の判定に係る検査法(ΔΔCq法)の開発と妥当性確認2023

    • Author(s)
      高畠令王奈、江木智宏、曽我慶介、峯岸恭孝、大西真理、成島純平、吉場聡子、柴田識人、中村公亮、近藤一成、岸根雅宏、真野潤一、橘田和美
    • Organizer
      日本食品化学会第29回総会・学術大会
    • Related Report
      2023 Annual Research Report
  • [Presentation] ゲノム編集イネにおけるin vitroオフターゲット予測法SITE-Seqを用いたオフターゲット予測性能の評価2023

    • Author(s)
      成島純平, 杉野御祐, 曽我慶介, 吉場聡子, 近藤一成, 柴田識人
    • Organizer
      日本ゲノム編集学会第7回大会
    • Related Report
      2023 Annual Research Report
  • [Presentation] 改正食品表示基準に対応した遺伝子組換えとうもろこし混入の判定に係る定性PCR検査法の開発2021

    • Author(s)
      曽我慶介, 中村公亮, 成島純平, 吉場聡子, 木俣真弥, 江木智宏, 岸根雅宏, 真野潤一, 橘田和美, 高畠令王奈, 柴田識人, 近藤一成
    • Organizer
      日本食品衛生学会第117会学術講演会
    • Related Report
      2021 Research-status Report
  • [Presentation] 改正食品表示基準に対応した遺伝子組換えダイズ混入の判定に係る定性PCR検査法の開発2021

    • Author(s)
      高畠令王奈, 江木智宏, 曽我慶介, 峯岸恭孝, 成島純平, 吉場聡子, 柴田識人, 中村公亮, 近藤一成, 岸根雅宏, 真野潤一, 橘田和美
    • Organizer
      日本食品衛生学会第117会学術講演会
    • Related Report
      2021 Research-status Report
  • [Presentation] ゲノム編集作物におけるオフターゲット予測法の検討2021

    • Author(s)
      成島純平, 木俣真弥, 志波優, 権藤崇裕, 秋元智, 曽我慶介, 吉場聡子, 中村公亮, 柴田識人, 近藤一成
    • Organizer
      第44回日本分子生物学会年会
    • Related Report
      2021 Research-status Report
  • [Presentation] 全ゲノム配列を用いた遺伝子変異検出におけるツール間比較2020

    • Author(s)
      曽我慶介、成島純平、吉場聡子、柴田識人、近藤一成
    • Organizer
      日本薬学会
    • Related Report
      2020 Research-status Report
  • [Presentation] ナノポアシーケンス技術を用いた致死性有毒キノコAmanita virosaのゲノムアセンブリの検討2020

    • Author(s)
      曽我慶介、吉田光範、坂田こずえ、近藤一成
    • Organizer
      日本分子生物学会
    • Related Report
      2020 Research-status Report

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Published: 2020-04-28   Modified: 2025-01-30  

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