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 were formed by microcell-mediated transfer of neo-tagged chromosome 8 into the cells derived from severe combined immunodeficiency (SCID) mouse. Thus, 110 cosmid markers were isolated from SV40-transformed SCID fibroblast cell line (SCVA) containing a p12-q11.1 region of human chromosome 8 and were assigned to eight regions in 8p12-q11.1, using a microcell-hybrid panel. For positional cloning of a human gene that restores the DNA-repair defect in a mouse with SCID on 8p11.1-q11.1 (SCID region), we constructed a yeast artificial chromosome (YAC) contig of about 4.5 Mb. Overlapping YACs were further aligned by restriction mapping, using rare-cutting restriction endonucleases. The cosmids and YAC contig should facilitate isolation of the SCID gene and other genes, such as the Werner syndrome-responsible gene in or near this region.
Department of Molecular and Cell Genetics, School of Life Science, Faculty of Medicine, Tottori University, Japan.
A
A
Kurimasa
N
N
Suzuki
S
S
Kumano
H
H
Li
D
D
Wells
M J
MJ
Wagner
F
F
Chen
D J
DJ
Chen
M
M
Oshimura
D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D002461:Cell Line, Transformed; D002874:Chromosome Mapping; D018244:Chromosomes, Artificial, Yeast; D002898:Chromosomes, Human, Pair 8; D003360:Cosmids; D005347:Fibroblasts; D018014:Gene Transfer Techniques; D005819:Genetic Markers; D006801:Humans; D006822:Hybrid Cells; D051379:Mice; D016513:Mice, SCID; D008969:Molecular Sequence Data
D005819:Genetic Markers