Generation and fluorescent in situ hybridization mapping of yeast artificial chromosomes of 1p, 17p, 17q, and 19q from a hybrid cell line by high-density screening of an amplified library

Generation and fluorescent in situ hybridization mapping of yeast artificial chromosomes of 1p, 17p, 17q, and 19q from a hybrid cell line by high-density screening of an amplified library

A yeast artificial chromosome (YAC) library has been constructed from a somatic cell hybrid containing a t(1p;19q) chromosome and chromosome 17. After amplification, part of this library was analyzed by high-density colony filter screening with a repetitive human DNA probe (Alu). The human YACs dist...

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Bibliographic Details
Published in:Genomics (San Diego, Calif.) Vol. 11; no. 4; p. 1079
Main Authors: Driesen, M S, Dauwerse, J G, Wapenaar, M C, Meershoek, E J, Mollevanger, P, Chen, K L, Fischbeck, K H, van Ommen, G J
Format: Journal Article
Language:English
Published: United States 01-12-1991
Subjects:
Chromosome Mapping
Chromosomes, Fungal
Chromosomes, Human, Pair 1
Chromosomes, Human, Pair 17
Chromosomes, Human, Pair 19
Cloning, Molecular
DNA Fingerprinting
Gene Amplification
Gene Library
Genome, Human
Humans
Hybrid Cells
Microscopy, Fluorescence
Nucleic Acid Hybridization
Polymerase Chain Reaction
Repetitive Sequences, Nucleic Acid
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Summary:A yeast artificial chromosome (YAC) library has been constructed from a somatic cell hybrid containing a t(1p;19q) chromosome and chromosome 17. After amplification, part of this library was analyzed by high-density colony filter screening with a repetitive human DNA probe (Alu). The human YACs distinguished by the screening were further analyzed by Alu fingerprinting and Alu PCR. Fluorescent in situ hybridization (FISH) was performed to localize the YACs to subchromosomal regions of chromosome 1p, 17, or 19q. We have obtained a panel of 123 individual YACs with a mean size of 160 kb, and 77 of these were regionally localized by FISH: 33 to 1p, 10 to 17p, 25 to 17q, and 9 to 19q. The YACs cover a total of 19.7 Mb or 9% of the 220 Mb of human DNA contained in the hybrid. No overlapping YACs have yet been detected. These YACs are available upon request and should be helpful in mapping studies of disease loci, e.g., Charcot-Marie-Tooth disease, Miller-Dieker syndrome, hereditary breast tumor, myotonic dystrophy, and malignant hyperthermia.
ISSN:0888-7543
DOI:10.1016/0888-7543(91)90035-D