Construction of a segmental chromosomal deletion in chromosomes II, III, IV, V and IX. (A) Illustration of each target chromosome, deleted region and hybridization probe location. Deleted regions are shown as black dotted lines on the left, right and internal side of each chromosome. Probes 1, 2, 3, 4, 5 and 6 correspond to coordinate number between 768 247–768 628 of chromosomes II, 17 312–17 705 of chromosome III, 1 517 154–1 517 535 of chromosome IV, 26 785–27 159 of chromosome V, 277 726–278 122 and 37 397–37 878 of chromosome IX, respectively. The length of each deleted region is indicated. The blue box, green box, red box, yellow circle and black circle represent the CgHIS3 marker, CgLEU2 marker, probe, artificial CEN4 and natural CEN sequence, respectively. (B) PFGE and Southern hybridization showing karyotype analysis of the parental haploid and transformant clones carrying segmental deletion on chromosomes II, III, IV, V and IX. For all chromosomes, probes were prepared by PCR amplification of a 400-bp internal sequence of the target region.