title: Construction of a 2.6-Mb contig in yeast artificial chromosomes spanning the human dystrophin gene using an STS-based approach.
abstract: A sequence tagged site (STS)-based approach has been used to construct a 2.6-Mb contig in yeast artificial chromosomes (YACs) spanning the human dystrophin gene. Twenty-seven STSs were used to identify and overlap 34 YAC clones. A DNA fingerprint of each clone produced by direct Alu-PCR amplification of YAC colonies and the isolation of YAC insert ends by vectorette PCR were used to detect overlaps in intron 1 (280 kb) where no DNA sequence data were available, thereby achieving closure of the map. This study has evaluated methods for mapping large regions of the X chromosome and provides a valuable resource of the dystrophin gene in cloned form for detailed analysis of gene structure and function in the future.
authors.affiliation: Paediatric Research Unit, United Medical School of Guy's Hospital, London, United Kingdom.
authors.forename: A J
authors.initials: AJ
authors.lastname: Coffey
authors.forename: R G
authors.initials: RG
authors.lastname: Roberts
authors.forename: E D
authors.initials: ED
authors.lastname: Green
authors.forename: C G
authors.initials: CG
authors.lastname: Cole
authors.forename: R
authors.initials: R
authors.lastname: Butler
authors.forename: R
authors.initials: R
authors.lastname: Anand
authors.forename: F
authors.initials: F
authors.lastname: Giannelli
authors.forename: D R
authors.initials: DR
authors.lastname: Bentley
mesh_terms: D001483:Base Sequence; D015139:Blotting, Southern; D015825:Chromosomes, Fungal; D003001:Cloning, Molecular / Q000379:methods; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D016172:DNA Fingerprinting; D015342:DNA Probes; D016189:Dystrophin / Q000235:genetics*; D015723:Gene Library; D015894:Genome, Human; D006801:Humans; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009838:Oligodeoxyribonucleotides; D016133:Polymerase Chain Reaction / Q000379:methods; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D016324:Sequence Tagged Sites
chemical_list: D015342:DNA Probes; D016189:Dystrophin; D009838:Oligodeoxyribonucleotides; D004247:DNA