Marker removal from actinomycetes genome using Flp recombinase.

Albert-Ludwigs-University of Freiburg, Pharmazeutische Biologie und Biotechnologie, Stefan-Meier st. 19, Freiburg 79104, Germany.

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.

PMID: 18550297 [PubMed - indexed for MEDLINE]