Targeted deletions created in yeast vectors by recombinational excision

We have developed a simple method for creating defined deletions in yeast vectors by utilizing the ability of Saccharomyces cerevisiae to perform homologous recombination. Two complementary single-stranded oligonucleotides are designed so that the 5' and 3' halves of the resulting double-s...

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Bibliographic Details
Published in:Nucleic acids research 1999-04, Vol.27 (8), p.e1-1
Main Authors: Dunø, M, Bendixen, C, Krejci, L, Thomsen, B
Format: Article
Language:English
Subjects:
DNA Restriction Enzymes - metabolism
Genetic Vectors
Oligonucleotides - chemistry
Plasmids
Recombination, Genetic
Saccharomyces cerevisiae - genetics
Sequence Deletion
Transformation, Genetic
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Summary:We have developed a simple method for creating defined deletions in yeast vectors by utilizing the ability of Saccharomyces cerevisiae to perform homologous recombination. Two complementary single-stranded oligonucleotides are designed so that the 5' and 3' halves of the resulting double-stranded oligonucleotide are homologous to the 5' and 3' side of a desired deletion junction, respectively. The sequence to be deleted is cleaved by restriction endonuclease digestion, followed by co-transformation of the linearized plasmid and the oligonucleotide into yeast. By homologous recombination in vivo, a subset of the plasmids will recircularize and simultaneously acquire the deletion as defined by the oligonucleotide.
ISSN:0305-1048
1362-4962
1362-4962
DOI:10.1093/nar/27.8.e1