Construction of a yeast artificial chromosome contig encompassing the human alpha 5(IV) collagen gene (COL4A5).
A PCR-based screening approach was used to isolate six yeast artificial chromosome (YAC) clones containing segments of the human alpha 5(IV) collagen gene (COL4A5). This gene is located at Xq22 and is known to be involved in the kidney disorder known as Alport syndrome (AS). By analyzing sequence-tagged sites, cDNA content, and rare-cutting restriction site patterns in these YAC clones, a contig that spans the entirety of the alpha 5(IV) gene was constructed. This contig may contain as much as 690 kb of DNA from the alpha 5(IV) locus. On the basis of the information obtained from these YAC clones, the genomic map and gene structure of the alpha 5(IV) gene have been refined. This study has also provided a valuable resource for subsequent studies of the alpha 5(IV) gene and its flanking DNA sequences.
Vetrie
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Paediatric Research Unit, United Medical School, Guy's Hospital, London, United Kingdom.
Flinter
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Bobrow
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Harris
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D001483:Base Sequence; D015139:Blotting, Southern; D015825:Chromosomes, Fungal; D003001:Cloning, Molecular; D003094:Collagen / Q000235:genetics*; D004277:DNA, Single-Stranded; D016521:Electrophoresis, Gel, Pulsed-Field; D015723:Gene Library; D015894:Genome, Human; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D015183:Restriction Mapping; D016324:Sequence Tagged Sites
D004277:DNA, Single-Stranded; D003094:Collagen