Genome replication, synthesis, and assembly of the bacteriophage T7 in a single cell-free reaction

ACS Synth Biol. 2012 Sep 21;1(9):408-13. doi: 10.1021/sb300049p. Epub 2012 Jul 10.

Abstract

The synthesis of living entities in the laboratory is a standing challenge that calls for innovative approaches. Using a cell-free transcription-translation system as a molecular programming platform, we show that the bacteriophage T7, encoded by a 40 kbp DNA program composed of about 60 genes, can be entirely synthesized from its genomic DNA in a test tube reaction. More than a billion infectious bacteriophages T7 per milliliter of reaction are produced after a few hours of incubation. The replication of the genomic DNA occurs concurrently with phage gene expression, protein synthesis, and viral assembly. The demonstration that genome-sized viral DNA can be expressed in a test tube, recapitulating the entire chain of information processing including the replication of the DNA instructions, opens new possibilities to program and to study complex biochemical systems in vitro.

Publication types

  • Letter
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Bacteriophage T7 / genetics*
  • Bacteriophage T7 / metabolism*
  • Cell-Free System*
  • DNA Replication*
  • DNA, Viral / genetics
  • Gene Expression
  • Genes, Viral

Substances

  • DNA, Viral