Different Thermostabilities of FLP and Cre Recombinases: Implications for Applied Site-Specific Recombination
Authors: Buchholz, Frank; Stewart, A. Francis
Source: Nucleic Acids Research, Volume 24, Number 21, November 1996 , pp. 4256-4262(7)
Publisher: Oxford University Press
Abstract:Genomic manipulations using site-specific recombinases rely on their applied characteristics in living systems. To understand their applied properties so that they can be optimally deployed, we compared the recombinases FLP and Cre in two assays. In both Escherichia coli and in vitro, FLP shows a different temperature optimum than Cre. FLP is more thermolabile, having an optimum near 30°C and little detectable activity above 39°C. Cre is optimally efficient at 37 °C and above. Consistent with FLP thermolability, recombination in a mammalian cell line mediated by a ligand- regulated FLP-androgen receptor fusion protein is more efficient at 35°C than at higher temperatures. We also document a mutation in a commercially available FLP plasmid (FLP-F70L) which renders this recombinase even more thermolabile. The different temperature optima of FLP, FLP-F70L and Cre influence their strategies of usage. Our results recommend the use of Cre for applications in mice that require efficient recombination. The thermolabilities of FLP and FLP-F70L can be usefully exploited for gain of function and cell culture applications.
Document Type: Research Article
Publication date: 1996-11-01
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