CONSTRUCTION OF 110 COSMID MARKERS AND A 4.5-MB YAC CONTIG ON HUMAN-CHROMOSOME 8P12-Q11

Microcell hybrids containing various regions of human chromosome 8 wer e formed by microcell-mediated transfer of neo-tagged chromosome 8 int o the cells derived from severe combined immunodeficiency (SCID) mouse . Thus, 110 cosmid markers were isolated from SV40-transformed SCID fi broblast cell line (SCVA) containing a p12-q11.1 region of human chrom osome 8 and were assigned to eight regions in 8p12-q11.1, using a micr ocell-hybrid panel. For positional cloning of a human gene that restor es the DNA-repair defect in a mouse with SCID on 8p11.1-q11.1 (SCID re gion), we constructed a yeast artificial chromosome (YAC) contig of ab out 4.5 Mb. Overlapping YACs were further aligned by restriction mappi ng, using rare-cutting restriction endonucleases. The cosmids and YAC contig should facilitate isolation of the SCID gene and other genes, s uch as the Werner syndrome-responsible gene in or near this region. (C ) 1995 Academic Press, Inc.