A series of conditional shuttle vectors for targeted genomic integration in budding yeast

Chia Ching Chou, Michael T. Patel, Marc R. Gartenberg

Research output: Contribution to journalArticlepeer-review

10 Scopus citations


The capacity of Saccharomyces cerevisiae to repair exposed DNA ends by homologous recombination has long been used by experimentalists to assemble plasmids from DNA fragments in vivo. While this approach works well for engineering extrachromosomal vectors, it is not well suited to the generation, recovery and reuse of integrative vectors. Here, we describe the creation of a series of conditional centromeric shuttle vectors, termed pXR vectors, that can be used for both plasmid assembly in vivo and targeted genomic integration. The defining feature of pXR vectors is that the DNA segment bearing the centromere and origin of replication, termed CEN/ARS, is flanked by a pair of loxP sites. Passaging the vectors through bacteria that express Cre recombinase reduces the loxP-CEN/ARS-loxP module to a single loxP site, thereby eliminating the ability to replicate autonomously in yeast. Each vector also contains a selectable marker gene, as well as a fragment of the HO locus, which permits targeted integration at a neutral genomic site. The pXR vectors provide a convenient and robust method to assemble DNAs for targeted genomic modifications.

Original languageEnglish (US)
Pages (from-to)1-9
Number of pages9
JournalFEMS Yeast Research
Issue number3
StatePublished - May 1 2015

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Applied Microbiology and Biotechnology


  • ARS
  • CEN
  • Cre
  • LoxP
  • Plasmid
  • Shuttle vector


Dive into the research topics of 'A series of conditional shuttle vectors for targeted genomic integration in budding yeast'. Together they form a unique fingerprint.

Cite this