A Nobel cloning tool using the designed horizontal gene-transfer system by natural transformation

• Project/Area Number: 23710239
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Applied genomics
• Research Institution: Tokyo Institute of Technology
• Principal Investigator: KANEKO Shinya 東京工業大学, 生命理工学研究科, 助教 (10399694)
• Project Period (FY): 2011 – 2013
• Project Status: Completed (Fiscal Year 2013)
• Budget Amount: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2013: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2012: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2011: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: 応用微生物 / 核酸 / ゲノム / バイオテクノロジー / 形質転換

Research Abstract

A designed horizontal gene-transfer system by natural transformation has been developed as novel cloning tool to introduce plasmid DNA into the objective cells without purification. Each culture condition for donor and recipient cells has been decided in this study. The bacteriolysis induction of donor cells by virulent phage is very important for universal use. Furthermore it was revealed that each aurintricarboxylic acid as nuclease inhibitor and exonuclease I as accelerator is effective for the DNA transfer by mixing it appropriately. In this condition it was found that the DNA transfer was extended to successful size increase up to 100 kb. The damage-free handling of DNA larger than 100 kb in test tubes is increasingly difficult. The designed transfer system named as Culture Mix Method (CMM) can apply various plasmid DNAs simply and conveniently, which should facilitate development of experimental tools to mobilize giant DNAs for genomic syntheses in synthetic biology.

Research Products

• [Presentation] 水平伝播を利用したクローニングシステム(CMM)の汎用性 A versatile designed horizontal gene-transfer system by natural transformation, named as Culture MixMethod (CMM) for genomic syntheses (2014)
  • Author(s): 金子真也、板谷光泰
  • Organizer: 日本農芸化学会2014年度(平成26年度)大会[東京]
  • Place of Presentation: 明治大学生田キャンパス(神奈川県川崎市)
  • Year and Date: 2014-03-30
• [Presentation] 細胞外核酸による水平伝播システム(CMM)を用いたゲノム合成 A designed horizontal gene-transfer system by natural transformation, named as Culture Mix Method (CMM) for genomic syntheses (2012)
  • Author(s): 金子真也、板谷光泰
  • Organizer: 第35回日本分子生物学会年会ワークショップ 3W3III-1セッションタイトル;「ゲノムを作り、種を越えて動かす : 合成生物学的観点から見る水平伝播」
  • Place of Presentation: 福岡国際会議場 4F小会議室 (福岡県)
  • Year and Date: 2012-12-13
  • Invited
• [Presentation] 細胞外核酸の水平伝播を介するゲノム合成 Genome synthesis via horizontal transfer of stable extra-cellular DNA (2011)
  • Author(s): 板谷光泰、金子真也
  • Organizer: 第84回日本生化学会大会シンポジウム「細胞外核酸の動態 : 医科学環境科学での新しい観点からの研究、応用展開」
  • Place of Presentation: 京都国際会館(京都府)
  • Year and Date: 2011-09-24
• [Presentation] 細胞外核酸の水平伝播を介するゲノム合成 Genome synthesis via horizontal transfer of stable extra-cellular DNA (2011)
  • Author(s): 板谷光泰、金子真也
  • Organizer: 第84回日本生化学会大会 シンポジウム「細胞外核酸の動態：医科学環境科学での新しい観点からの研究、応用展開」
  • Place of Presentation: 京都国際会館(京都府)
• [Presentation] 水平伝播を利用したクローニングシステム(CMM)の汎用性 A versatile designed horizontal gene-transfer system by natural transformation, named as Culture Mix Method (CMM) for genomic syntheses.
  • Author(s): 金子真也、板谷光泰
  • Organizer: 日本農芸化学会2014年度(平成26年度)大会[東京]
  • Place of Presentation: 明治大学生田キャンパス (神奈川県川崎市)
• [Presentation] 細胞外核酸による水平伝播システム(CMM)を用いたゲノム合成 A designed horizontal gene-transfer system by natural transformation, named as Culture Mix Method (CMM) for genomic syntheses.
  • Author(s): 金子真也、板谷光泰
  • Organizer: 第35回日本分子生物学会年会 ワークショップ 3W3III-1 セッションタイトル；「ゲノムを作り、種を越えて動かす：合成生物学的観点から見る水平伝播」
  • Place of Presentation: 福岡国際会議場 4F 小会議室 (福岡県)
  • Invited