Identification of orthogonal LoxPsym sites for controlled recombination in S. cerevisiae, Z. mays and E. coli
We developed a set of novel LoxPsym sites. We evaluated orthogonality of these sites in S. cerevisiae, Z. mays and E. coli. For experiments in S. cerevisiae, we used a fluorescent reporter and Sanger sequencing to identify the structural rearrangements in an array of LoxPsym variants after induction of recombination. In Z. mays, we used a library of plasmids with pairwise combinations of LoxPsym sites and determined the abundance of recombination for each LoxPsym pair using next generation sequencing. In E.coli, we used a PCR-based approach and sequenced recombined sites by Sanger sequencing.