PCR-directed in vivo plasmid construction using homologous recombination in baker's yeast.

Erik C. Andersen*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

6 Scopus citations

Abstract

A variety of applications require the creation of custom-designed plasmids, including transgenic reporters, heterologous gene fusions, and phenotypic rescue plasmids. These plasmids are created traditionally using restriction digests and in vitro ligation reactions, but these techniques are dependent on available restriction sites and can be laborious given the size and number of fragments to be ligated. The baker's yeast Saccharomyces cerevisiae provides a powerful platform to create nearly any plasmid through PCR-directed yeast-mediated ligation. This technique can ligate complex plasmids of up to 50 kilobasepairs (kb) in vivo to produce plasmids with precisely defined sequences.

Original languageEnglish (US)
Pages (from-to)409-421
Number of pages13
JournalMethods in molecular biology (Clifton, N.J.)
Volume772
StatePublished - 2011

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Fingerprint Dive into the research topics of 'PCR-directed in vivo plasmid construction using homologous recombination in baker's yeast.'. Together they form a unique fingerprint.

Cite this