RAPID AND EFFICIENT CONSTRUCTION OF YEAST ARTIFICIAL CHROMOSOME CONTIGS IN THE MOUSE GENOME WITH INTERSPERSED REPETITIVE SEQUENCE PCR (IRS-PCR) - GENERATION OF A 5-CM, GREATER-THAN-5 MEGABASE CONTIG ON MOUSE CHROMOSOME-1

We have developed a new technique for the generation of YAC contigs in the mouse genome that is based on the ability to detect overlapping clones by hybridization of shared IRS-PCR products. As a demonstration of the technique, a 5-cM, >5 megabase contig was developed on the distal half of mouse Chromosome (Chr) 1, spanning the region from Lamb2 to At3. The contig covers roughly 5% of the genetic distance of the chromosome and is comprised of more than 80 clones; 71 probes were assigned physical order on the chromosome, of which 59 were new markers generated in this study. Eight of the new probes were shown to be polymorphic between C3H/HeJ-gld and M. spretus. Three probes were mapped on a [(C3H/HeJ-gld X M. spretus) X C3H/HeJ-gld] interspecific backcross to integrate the physical map with a high-resolution genetic map of the region.