Functional expression of the Cre recombinase in actinomycetes.
Site-specific recombinases revolutionized "in vivo" genetic engineering because they can catalyze precise excisions, integrations, inversions, or translocations of DNA between their distinct recognition target sites. We have constructed a synthetic gene encoding Cre recombinase with the GC content 67.7% optimized for expression in high-GC bacteria and demonstrated this gene to be functional in Streptomyces lividans. Using the synthetic cre(a) gene, we have removed an apramycin resistance gene flanked by loxP sites from the chromosome of S. lividans with 100% efficiency. Sequencing of the chromosomal DNA part showed that excision of the apramycin cassette by Cre recombinase was specific.
Fedoryshyn
Marta
M
Albert-Ludwigs-Universität, Institut für Pharmazeutische Wissenschaften, Freiburg, Germany.
Welle
Elisabeth
E
Bechthold
Andreas
A
Luzhetskyy
Andriy
A
D003001:Cloning, Molecular; D015870:Gene Expression; D005818:Genetic Engineering / Q000379:methods*; D019426:Integrases / Q000235:genetics* / Q000378:metabolism; D010957:Plasmids / Q000235:genetics; D048170:Streptomyces lividans / Q000235:genetics* / Q000378:metabolism; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism
D014764:Viral Proteins; C045073:Cre recombinase; D019426:Integrases