Mapping of 386 kb of genomic DNA in the human dystrophin-encoding gene (DYS) using an ordered phage lambda sublibrary of a YAC clone containing the DYS region.
An integrated restriction map for HindIII and EcoRI has been constructed for 386 kb of the human dystrophin-encoding gene by partial digest mapping of 35 overlapping lambda EMBL3cosW phage clones derived from a yeast artificial chromosome containing this region. Map construction was simplified in two ways. Firstly, the sequence arrangement of lambda EMBL3cosW is such that only map data from cloned inserts are generated using partial digests of lambda phage DNA asymmetrically labelled at the left cos end with a complementary 32P-labelled oligodeoxyribonucleotide. Secondly, the degree of partial digestion was standardised for each restriction enzyme by using ultraviolet light-induced formation of thymine dimers in the recognition sequence to partially block the cleavage reaction. The map provides the basis for work on the analysis of chromosomal rearrangements in this region which give rise to Duchenne muscular dystrophy, and for studies of chromosome structure and function.
Wood
L
L
University Clinical Biochemistry, Southampton General Hospital, UK.
Whittaker
P A
PA
D000818:Animals; D010582:Bacteriophage lambda; D018244:Chromosomes, Artificial, Yeast; D003001:Cloning, Molecular / Q000379:methods; D015246:Deoxyribonuclease EcoRI; D015247:Deoxyribonuclease HindIII; D016189:Dystrophin / Q000235:genetics*; D015723:Gene Library; D015186:Hominidae / Q000235:genetics*; D006801:Humans; D009132:Muscles / Q000378:metabolism; D009838:Oligodeoxyribonucleotides; D011401:Promoter Regions, Genetic; D015183:Restriction Mapping
D016189:Dystrophin; D009838:Oligodeoxyribonucleotides; D015246:Deoxyribonuclease EcoRI; D015247:Deoxyribonuclease HindIII