DNA fragment transplacement in Saccharomyces cerevisiae: some genetic considerations.
The ability to make specific genomic alterations is an invaluable tool to researchers who use genetics and biochemistry to study problems in biology. We have investigated some of the parameters governing DNA fragment transplacement in two commonly used strains of Saccharomyces cerevisiae, S288C and W303-1A. These strains exhibited a marked difference in their capacity to take up plasmid DNA and utilize linear DNA fragments as substrates for transplacement. The contributions of transformation efficiency, length of homology, and alternative target site configuration were assessed. This analysis indicates that several genetic parameters are important for optimizing the efficiency of gene transplacement.
Manthey
Glenn M
GM
Division of Molecular Biology, Beckman Research Institute, City of Hope National Medical Center, Duarte, CA, USA.
Navarro
Michelle S
MS
Bailis
Adam M
AM
D001483:Base Sequence; D002455:Cell Division; D017931:DNA Primers; D004271:DNA, Fungal / Q000235:genetics*; D018274:Electroporation / Q000379:methods; D005821:Genetic Techniques; D005838:Genotype; D007202:Indicators and Reagents; D008969:Molecular Sequence Data; D016296:Mutagenesis; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods; D012441:Saccharomyces cerevisiae / Q000145:classification / Q000166:cytology / Q000235:genetics*; D014170:Transformation, Genetic
D017931:DNA Primers; D004271:DNA, Fungal; D007202:Indicators and Reagents