Applications of the Saccharomyces cerevisiae Flp-FRT System in Bacterial GeneticsSchweizer H.P.
Department of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, Colo., USA
The Flp-FRT site-specific recombinationsystem from Saccharomyces cerevisiae is a powerful and efficient tool for high-throughput genetic analysis of bacteria in the postgenomic era. This review highlights the features of the Flp-FRT system, describes current bacterial genetic methods incorporating this technology and, finally, suggests potential future uses of this system. In combination with improved allele replacement methods, recyclable FRT mutagenesis cassettes, whose antibiotic resistance markers can be excised from the chromosome in vivo, are useful for the rapid construction of multiple, unmarked mutations in the same chromosome, and thus aid in the generation of live vaccine strains or food-safe bacteria. The high-specificity of the Flp-FRT system makes it also applicable for manipulation of whole genomes, including in vivo cloning of large genomic segments. Integration-proficient vectors, from which antibiotic resistance markers and replication functions can be evicted after integration of the desired sequences into the chromosome, are useful for the construction of strains destined for environmental release, e.g. strains used as biosensors or for bioremediation. Although the Flp-FRT system is extremely efficient and easy to use, its true potential in bacterial genetics has not yet been fully exploited. On the contrary, in many instances this technology is probably greatly underutilized, especially in gram-positive bacteria.
© 2003 S. Karger AG, Basel
Dr. Herbert P. Schweizer
Department of Microbiology, Immunology and Pathology
Colorado State University
Fort Collins, CO 80523-1682A (USA)
Tel. +1 970 491 3536, Fax +1 970 491 1815, E-Mail Herbert.Schweizer@colostate.edu
Number of Print Pages : 11
Number of Figures : 4, Number of Tables : 0, Number of References : 76
Journal of Molecular Microbiology and Biotechnology
Founded 1999 by M.H. Saier, Jr.
Vol. 5, No. 2, Year 2003 (Cover Date: Released May 2003)
Journal Editor: M.H. Saier, Jr., La Jolla, Calif.
ISSN: 1464–1801 (print), 1660–2412 (Online)
For additional information: http://www.karger.com/mmb