Figure 1. Recombination system for the insertion of the R403stop mutation into the mutase locus of pBAC_MMA.

The I-Sce I/KanR cassette is flanked with 50 bp regions which are homologous to sequences 50 bp on either side of the site to be targeted in exon 6, and was utilized to introduce the cassette in the first stage. A 255 bp PCR product containing the R403stop mutation was used in the second stage to replace the I-Sce I/KanR cassette and produce pBAC_MMA*, R403stop.