Designer deletion strains derived from Saccharomyces cerevisiae S288C: a useful set of strains and plasmids for PCR-mediated gene disruption and other applications.
A set of yeast strains based on Saccharomyces cerevisiae S288C in which commonly used selectable marker genes are deleted by design based on the yeast genome sequence has been constructed and analysed. These strains minimize or eliminate the homology to the corresponding marker genes in commonly used vectors without significantly affecting adjacent gene expression. Because the homology between commonly used auxotrophic marker gene segments and genomic sequences has been largely or completely abolished, these strains will also reduce plasmid integration events which can interfere with a wide variety of molecular genetic applications. We also report the construction of new members of the pRS400 series of vectors, containing the kanMX, ADE2 and MET15 genes.
Brachmann
C B
CB
Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
Davies
A
A
Cost
G J
GJ
Caputo
E
E
Li
J
J
Hieter
P
P
Boeke
J D
JD
D000483:Alleles; D015152:Blotting, Northern; D015139:Blotting, Southern; D017931:DNA Primers / Q000737:chemistry; D017353:Gene Deletion; D015870:Gene Expression; D005819:Genetic Markers / Q000502:physiology*; D016681:Genome, Fungal; D009154:Mutation / Q000502:physiology*; D010957:Plasmids / Q000737:chemistry* / Q000235:genetics / Q000502:physiology; D016133:Polymerase Chain Reaction; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000502:physiology
D017931:DNA Primers; D005819:Genetic Markers