Marker removal from actinomycetes genome using Flp recombinase.

Marta Fedoryshyn, Lutz Petzke, Elisabeth Welle, Andreas Bechthold, Andriy Luzhetskyy, Marta Fedoryshyn, Lutz Petzke, Elisabeth Welle, Andreas Bechthold, Andriy Luzhetskyy, Marta Fedoryshyn, Lutz Petzke, Elisabeth Welle, Andreas Bechthold, Andriy Luzhetskyy

Index: Gene 419(1-2) , 43-7, (2008)

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Abstract

We report here a system for the functional expression of the Flp recombinase in several actinomycetes: Streptomyces coelicolor, S. lividans, and Saccharotrix espanaensis. We have constructed a synthetic gene encoding the Flp recombinase with a GC content of 60.6% optimized for expression in high-GC bacteria. Using the synthetic flp(a) gene, we have removed an apramycin resistance gene flanked by FRT sites from the chromosome of actinomycetes with an efficiency of 40%. Sequencing the region of chromosome showed that excision of the apramycin cassette by Flp recombinase was specific.