Cell-to-cell direct cloning by designed horizontal transfer using extra-cellular DNA

2016 Fiscal Year Final Research Report

• Project/Area Number: 26430197
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: System genome science
• Research Institution: Tokyo Institute of Technology
• Principal Investigator: KANEKO Shinya 東京工業大学, 生命理工学院, 助教 (10399694)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Keywords: ダイレクトクローニング / 大腸菌 / 枯草菌 / 応用微生物 / 形質転換 / 核酸 / ゲノム / バイオテクノロジー

Outline of Final Research Achievements

Cell-to-cell direct cloning technology has established by designed horizontal transfer using extra-cellular DNA without purified DNAs. The purified DNAs in a test tube have been generally required to introduce into the host cell for molecular cloning technology in laboratory. In the natural environment, extra-cellular DNAs without purified step contribute to the gene delivery during horizontal gene transfer (HGT) dependent on the stability of DNA released from donor cells. Under such a concept we have developed that DNA delivery procedure using lysate of Escherichia coli lysed by virulent bacteriophage into competent Bacillus subtilis cells. The direct cloning technology has been applied for the DNA synthesis by fragment assembly within the B. subtilis genome. Furthermore we showed the transfer of plasmid DNAs from lysing E. coli into cyanobacteria, which is suggests the procedure is useful for general use.

Free Research Field

応用ゲノム科学