Abstract
We have extended the technique of PCR-directed recombination in Saccharomyces cerevisiae to develop a simple method for plasmid or gene construction in the absence of suitable restriction sites. The DNA to be cloned is PCR-amplified with 30-40 bp of homology to a linearized yeast plasmid. Co-transformation into yeast results in homologous recombination at a position directed by the PCR oligonucleotides.
Publication types
-
Research Support, U.S. Gov't, P.H.S.
MeSH terms
-
Cloning, Molecular / methods
-
DNA Primers
-
DNA, Recombinant / genetics
-
Genes, Fungal
-
Genes, Reporter / genetics
-
Green Fluorescent Proteins
-
Luminescent Proteins / genetics
-
Luminescent Proteins / metabolism
-
Plasmids / genetics
-
Plasmids / metabolism*
-
Polymerase Chain Reaction
-
Recombination, Genetic / genetics*
-
Saccharomyces cerevisiae / genetics*
-
Transformation, Genetic / genetics
Substances
-
DNA Primers
-
DNA, Recombinant
-
Luminescent Proteins
-
Green Fluorescent Proteins