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  • Self-replication of DNA by its encoded proteins in liposome-based synthetic cells.

Self-replication of DNA by its encoded proteins in liposome-based synthetic cells.

Nature communications (2018-04-22)
Pauline van Nies, Ilja Westerlaken, Duco Blanken, Margarita Salas, Mario Mencía, Christophe Danelon
ABSTRACT

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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Acridine Orange hydrochloride solution, 10 mg/mL in H2O, ≥95.0% (HPLC)
Sigma-Aldrich
L-Rhamnose, natural sourced, 99%, FG