Construction of a 2.6-Mb contig in yeast artificial chromosomes spanning the human dystrophin gene using an STS-based approach.
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.
Coffey
A J
AJ
Paediatric Research Unit, United Medical School of Guy's Hospital, London, United Kingdom.
Roberts
R G
RG
Green
E D
ED
Cole
C G
CG
Butler
R
R
Anand
R
R
Giannelli
F
F
Bentley
D R
DR
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
D015342:DNA Probes; D016189:Dystrophin; D009838:Oligodeoxyribonucleotides; D004247:DNA