Molecular and General Genetics MGG

, Volume 219, Issue 1, pp 320–323

Use of site-specific recombination to regenerate selectable markers

Authors

  • James M. Cregg
    • The Salk Institute Biotechnology/Industrial Associates, Inc.
  • Knut R. Madden
    • The Salk Institute Biotechnology/Industrial Associates, Inc.
Short Communications

DOI: 10.1007/BF00261194

Cite this article as:
Cregg, J.M. & Madden, K.R. Molec. Gen. Genet. (1989) 219: 320. doi:10.1007/BF00261194

Summary

A method which allows the repeated use of a single selectable marker in DNA transformations was demonstrated. This marker regeneration method employed portions of the Saccharomyces cerevisiae 2 μm circle plasmid: the inverted repeat sequences (FRTs), and the FLP gene whose product, a site-specific recombinase, catalyzes recombination events between FRTs. When FRTs were oriented as direct repeats and integrated into the genome of the yeast Pichia pastoris, FLP-mediated recombination resulted in the efficient and precise deletion of DNA located between the repeats. In the example described, the S. cerevisiae ARG4 gene, placed between a set of FRTs and integrated into Pichia in a prior transformation, was deleted by FLP, thereby regenerating an arginine-requiring phenotype in the P. pastoris strain.

Key words

2 μm plasmidFLP genePichia pastorisMethylotrophic yeast

Copyright information

© Springer-Verlag 1989