Yeast artificial chromosomes spanning 8 megabases and 10-15 centimorgans of human cytogenetic band Xq26.
A successful test is reported to generate long-range contiguous coverage of DNA from a human cytogenetic band in overlapping yeast artificial chromosomes (YACs). Seed YACs in band Xq26 were recovered from a targeted library of clones from Xq24-q28 with 14 probes, including probes for the hypoxanthine guanine phosphoribosyltransferase- and coagulation factor IX-encoding genes and nine probes used in linkage mapping. Neighboring YACs were then identified by 25 "walking" steps with end-clones, and the content of 71 probes in cognate YACs was verified by further hybridization analyses. The resultant contig extends across 8 million base pairs, including most of band Xq26, with an order of markers consistent with linkage data. YAC-based mapping, thus, permits steps toward a fully integrated physical and genetic map and is probably adequate to sustain most of the human genome project.
Little
R D
RD
Department of Molecular Microbiology, Washington University School of Medicine, St. Louis, MO 63110.
Pilia
G
G
Johnson
S
S
D'Urso
M
M
Schlessinger
D
D
D015139:Blotting, Southern; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D005796:Genes; D005819:Genetic Markers; D005822:Genetic Vectors; D006801:Humans; D009693:Nucleic Acid Hybridization; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae; D014960:X Chromosome / Q000648:ultrastructure*
D005819:Genetic Markers