A fully decompressed synthetic bacteriophage øX174 genome assembled and archived in yeast

Virology. 2012 Dec 20;434(2):278-84. doi: 10.1016/j.virol.2012.09.020. Epub 2012 Oct 15.


The 5386 nucleotide bacteriophage øX174 genome has a complicated architecture that encodes 11 gene products via overlapping protein coding sequences spanning multiple reading frames. We designed a 6302 nucleotide synthetic surrogate, øX174.1, that fully separates all primary phage protein coding sequences along with cognate translation control elements. To specify øX174.1f, a decompressed genome the same length as wild type, we truncated the gene F coding sequence. We synthesized DNA encoding fragments of øX174.1f and used a combination of in vitro- and yeast-based assembly to produce yeast vectors encoding natural or designer bacteriophage genomes. We isolated clonal preparations of yeast plasmid DNA and transfected E. coli C strains. We recovered viable øX174 particles containing the øX174.1f genome from E. coli C strains that independently express full-length gene F. We expect that yeast can serve as a genomic 'drydock' within which to maintain and manipulate clonal lineages of other obligate lytic phage.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Bacteriophage phi X 174 / genetics*
  • DNA, Viral / genetics*
  • Escherichia coli / genetics
  • Escherichia coli / virology
  • Genetic Engineering / methods*
  • Genome, Viral*
  • Molecular Biology / methods
  • Plasmids
  • Saccharomyces cerevisiae / genetics


  • DNA, Viral