<i>In vitro</i> Reconstitution of Replication Cycle of the <i>Escherichia coli</i> Chromosome and its Impact on Synthetic Biology

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  • 大腸菌染色体複製サイクルの試験管内再構成と合成生物学
  • ダイチョウキン センショクタイ フクセイ サイクル ノ シケンカン ナイ サイコウセイ ト ゴウセイ セイブツガク

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Abstract

<p>In vitro reconstitution of cellular events is a promising strategy as a bottom-up approach to build an artificial cell in the field of synthetic biology. Also, the reconstituted system sometimes provides a useful biotechnology tool for various fields. We recently reconstituted a whole replication cycle of the Escherichia coli chromosome in vitro as a part of the bottom-up synthetic biology. This replication cycle reaction (RCR) propagates circular DNA exponentially and precisely by autonomous repetition of the cycle in an isothermal reaction. Even genome-sized DNA (1 Mbp) can be propagated as circular DNA in RCR. We also developed a DNA assembly tool. The combination of the DNA assembly and RCR amplification enables cell-free synthesis of artificial large circular DNA, providing a powerful tool for a top-down study of the synthetic biology, in which living cells are modified at a whole genome level.</p>

Journal

  • Seibutsu Butsuri

    Seibutsu Butsuri 60 (5), 284-287, 2020

    The Biophysical Society of Japan General Incorporated Association

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