Back to Search Start Over

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

Authors :
Jaschke, Paul R.
Lieberman, Erica K.
Rodriguez, Jon
Sierra, Adrian
Endy, Drew
Source :
Virology. 12/20/2012, Vol. 434 Issue 2, p278-284. 7p.
Publication Year :
2012

Abstract

Abstract: 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. [Copyright &y& Elsevier]

Details

Language :
English
ISSN :
00426822
Volume :
434
Issue :
2
Database :
Academic Search Index
Journal :
Virology
Publication Type :
Academic Journal
Accession number :
84186290
Full Text :
https://doi.org/10.1016/j.virol.2012.09.020