Fig. 1

Overview of replacement analysis of target region. Target region was replaced by DNA module harboring CgLEU2. For amplification of DNA module, forward and reverse primers were designed to anneal with the plasmid pSJ69 and DNA module was amplified by PCR. DNA module has 50 bp homology sequence with the target region (harboring A1, A2, A2 (Ex), B1, B1 (Ex) and B2 sub-regions) in both edges. After introduction of DNA module by transformation into yeast cell, target region was supposed to be replaced by the DNA module through homologous recombination. Replacement of left edge of target region was checked by colony PCR with specific forward primers depending upon the target regions and common reverse primer SJP 119 which leads to PCR product of 1 kb. On the other hand, replacement of right edge of target region was checked with common forward primer SJP 127 and specific reverse primers depending upon respective regions which also amplified 1 kb band