Marker removal from actinomycetes genome using Flp recombinase.
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.
Albert-Ludwigs-University of Freiburg, Pharmazeutische Biologie und Biotechnologie, Stefan-Meier st. 19, Freiburg 79104, Germany.
Marta
M
Fedoryshyn
Lutz
L
Petzke
Elisabeth
E
Welle
Andreas
A
Bechthold
Andriy
A
Luzhetskyy
D000192:Actinomycetales / Q000235:genetics*; D015825:Chromosomes, Fungal; D004254:DNA Nucleotidyltransferases / Q000235:genetics / Q000378:metabolism*; D004352:Drug Resistance, Microbial / Q000235:genetics*; D015870:Gene Expression; D005813:Genes, Synthetic; D005818:Genetic Engineering / Q000379:methods*; D005819:Genetic Markers; D016681:Genome, Fungal; D009328:Nebramycin / Q000031:analogs & derivatives / Q000494:pharmacology; D011995:Recombination, Genetic; D048372:Streptomyces coelicolor / Q000235:genetics; D048170:Streptomyces lividans / Q000235:genetics
D005819:Genetic Markers; D009328:Nebramycin; C011666:apramycin; D004254:DNA Nucleotidyltransferases; C045119:FLP recombinase