Generation of ordered phage sublibraries of YAC clones: Construction of a 400-kb phage contig in the human dystrophin gene

Journal Article · Mon Feb 01 04:00:00 EST 1993 · Genomics; (United States)

DOI:https://doi.org/10.1006/geno.1993.1089· OSTI ID:6660052

Whittaker, P A; Wood, L; Mathrubutham, M ^[1]; Anand, R ^[2]

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1. Southampton General Hospital (United Kingdom)
2. ICI Pharmaceuticals, Macclesfield Cheshire (United Kingdom)

A phage contig of 400 kb that extends from the brain-specific promoter at the 5[prime]-end of the human dystrophin gene, through the muscle-specific promoter over 100 kb further downstream, and across most of intron 1 has been assembled. To achieve this, a yeast artificial chromosome (YAC) subcloning approach was used. Total DNA from a yeast strain containing a 400-kb YAC from the dystrophin gene was cloned using [lambda] phage vector containing RNA polymerase promoters flanking the cloning sites. Phage containing human DNA inserts were then ordered into an overlapping set by hybridization of end-specific RNA probes from individual clones back to plaque lifts of gridded phage subclones. The clones generated will be useful as reagents for detailed structural and functional analyses of this region of the dystrophin gene. 6 refs., 2 figs.

• OSTI ID: 6660052
• Journal Information: Genomics; (United States), Journal Name: Genomics; (United States) Vol. 15:2; ISSN GNMCEP; ISSN 0888-7543
• Country of Publication: United States
• Language: English

Related Subjects

550400* -- Genetics
59 BASIC BIOLOGICAL SCIENCES
CLONING
DNA
GENES
HYBRIDIZATION
LIBRARIES
MOLECULAR STRUCTURE
NUCLEIC ACIDS
ORGANIC COMPOUNDS
PROBES
PROMOTERS
REAGENTS
RNA
SYNTHESIS