Chromosomal region of the cystic fibrosis gene in yeast artificial chromosomes: a model for human genome mapping.
A general strategy for cloning and mapping large regions of human DNA with yeast artificial chromosomes (YAC's) is described. It relies on the use of the polymerase chain reaction to detect DNA landmarks called sequence-tagged sites (STS's) within YAC clones. The method was applied to the region of human chromosome 7 containing the cystic fibrosis (CF) gene. Thirty YAC clones from this region were analyzed, and a contig map that spans more than 1,500,000 base pairs was assembled. Individual YAC's as large as 790 kilobase pairs and containing the entire CF gene were constructed in vivo by meiotic recombination in yeast between pairs of overlapping YAC's.
Green
E D
ED
Department of Genetics, Washington University School of Medicine, St. Louis, MO 63110.
Olson
M V
MV
D001483:Base Sequence; D002874:Chromosome Mapping; D015825:Chromosomes, Fungal; D002897:Chromosomes, Human, Pair 7; D003001:Cloning, Molecular / Q000379:methods*; D003550:Cystic Fibrosis / Q000235:genetics*; D004247:DNA / Q000235:genetics; D015894:Genome, Human; D006801:Humans; D008957:Models, Genetic; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D016133:Polymerase Chain Reaction / Q000379:methods; D011995:Recombination, Genetic; D012441:Saccharomyces cerevisiae / Q000235:genetics*
D015345:Oligonucleotide Probes; D004247:DNA