Vertical integration of cosmid and YAC resources for interval mapping on the X-chromosome.
The vertical integration of cosmid and yeast artificial chromosome (YAC) resources is of particular importance in the development of high-resolution maps of selected regions of the human genome. A resource of approximately 95,000 cosmids constructed using DNA from primary fibroblasts of karyotype 49,XXXXX was validated by detailed characterization of a 200-kb cosmid contig spanning exons 8-20 of the dystrophin gene. This resource was used to construct contigs in 0.65 Mb of Xq26 by hybridization of gel-purified YAC DNA to high-density gridded arrays of the cosmid library; positive cosmids were overlapped by finger-printing. Contigs were oriented and ordered relative to existing YACs in the region using cross-hybridization. The overlaps between a representative set of cosmids define 54 intervals of 5-20 kb and were used to construct a high-resolution cosmid interval map of the region, locating markers, dinucleotide repeats, and candidate CpG islands. This approach can be applied rapidly to large regions of the genome and without recourse to subcloning of individual YACs.
Holland
J
J
Paediatric Research Unit, United Medical School of Guy's Hospital, London, United Kingdom.
Coffey
A J
AJ
Giannelli
F
F
Bentley
D R
DR
D000782:Aneuploidy; D002460:Cell Line; D002874:Chromosome Mapping / Q000379:methods*; D015825:Chromosomes, Fungal; D003360:Cosmids; D004247:DNA; D015226:Dinucleoside Phosphates / Q000032:analysis; D015894:Genome, Human; D015698:Genomic Library; D006801:Humans; D007621:Karyotyping; D014960:X Chromosome
D004247:DNA; D015226:Dinucleoside Phosphates; C015772:CpG dinucleotide