Yeast-gene replacement using PCR products

Megan Bergkessel, Christine Guthrie (Lead / Corresponding author), John Abelson

Research output: Chapter in Book/Report/Conference proceedingChapter

2 Citations (Scopus)


It is often useful to replace a small region of the yeast genome containing a gene of interest with a selectable marker. The selectable marker allows for easy identification of yeast cells that have successfully carried out the gene replacement, and functional consequences of the loss of that gene can then be assessed. The same technique can also be used for removing noncoding portions of the genome that may be of interest, such as promoters or 3′ or 5′ UTRs, and for introducing tags on the N- or C-termini of proteins (alternatively, see a marker-free yeast gene replacement method on Gene Knockouts, in vivo site-directed mutagenesis and Other Modifications Using the Delitto Perfetto System in Saccharomyces cerevisiae).

Original languageEnglish
Title of host publicationLaboratory Methods in Enzymology
Subtitle of host publicationCell, Lipid and Carbohydrate
EditorsJon Lorsch
PublisherAcademic Press Inc.
Number of pages13
ISBN (Print)9780124200678
Publication statusPublished - 2013

Publication series

NameMethods in Enzymology
ISSN (Print)0076-6879
ISSN (Electronic)1557-7988


  • Agarose gel visualization
  • Colonies growing on selective media
  • Deoxyribonucleic acid (DNA)
  • Polyacrylamide gel electrophoresis (PAGE)
  • Polymerase chain reaction (PCR)
  • Transforming yeast with linear PCR product
  • Yeast-gene replacement

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology


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