Department of Bionanoscience, Kavli Institute of Nanoscience, Delft University of Technology, van der Maasweg 9, Delft, 2629 HZ, The Netherlands.
Centro de Biología Molecular "Severo Ochoa" (CSIC-UAM), Universidad Autónoma, Canto Blanco, Madrid, 28049, Spain.
Nat Commun. 2018 Apr 20;9(1):1583. doi: 10.1038/s41467-018-03926-1.
Replication of DNA-encoded information and its conversion into functional proteins are universal properties of life. In an effort toward the construction of a synthetic minimal cell, we implement here the DNA replication machinery of the Φ29 virus in a cell-free gene expression system. Amplification of a linear DNA template by self-encoded, de novo synthesized Φ29 proteins is demonstrated. Complete information transfer is confirmed as the copied DNA can serve as a functional template for gene expression, which can be seen as an autocatalytic DNA replication cycle. These results show how the central dogma of molecular biology can be reconstituted and form a cycle in vitro. Finally, coupled DNA replication and gene expression is compartmentalized inside phospholipid vesicles providing the chassis for evolving functions in a prospective synthetic cell relying on the extant biology.
DNA 编码信息的复制及其转化为功能性蛋白质是生命的普遍特性。在构建合成最小细胞的努力中，我们在这里在无细胞基因表达系统中实现了 Φ29 病毒的 DNA 复制机制。证明了通过自我编码、从头合成的 Φ29 蛋白对线性 DNA 模板的扩增。完整的信息传递得到了确认，因为复制的 DNA 可以作为基因表达的功能性模板，这可以看作是一个自我催化的 DNA 复制循环。这些结果表明，分子生物学的中心法则如何在体外重新构建并形成一个循环。最后，将 DNA 复制和基因表达耦联在磷脂囊泡内部，为在依赖现有生物学的有前景的合成细胞中进化功能提供了底盘。
