Jaschke PR, et al. (2012) A fully decompressed synthetic bacteriophage ?X174 genome assembled and archived in yeast. Virology 434(2):278-84
Abstract: The 5386 nucleotide bacteriophage ?X174 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, ?X174.1, that fully separates all primary phage protein coding sequences along with cognate translation control elements. To specify ?X174.1f, a decompressed genome the same length as wild type, we truncated the gene F coding sequence. We synthesized DNA encoding fragments of ?X174.1f 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 ?X174 particles containing the ?X174.1f 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.
|Status: Published||Type: Journal Article||PubMed ID: 23079106|
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