Cloning human telomeric DNA fragments into Saccharomyces cerevisiae using a yeast-artificial-chromosome vector
- Washington Univ. School of Medicine, St. Louis, MO (USA)
- Univ. of California, Los Alamos, NM (USA)
Telomeric fragments of human DNA ranging in size from 50 to 250 kilobases were cloned into Saccharomyces cerevisiae using a yeast-artificial-chromosome (YAC) vector. Six human-telomeric YAC (HTY) strains were selected by virtue of the specific hybridization of their DNA with the human telomeric terminal-repeat sequence (TTAGGG){sub n}, and the telomeric localization of this sequence within each YAC was demonstrated by its sensitivity to nuclease BAL-31. In situ hybridization of DNA from three of these HTY strains with human metaphase chromosomes yielded discrete patterns of hybridization signals at the telomeres of a limited number of human chromosomes, different for each clone. DNA from selected cosmid subclones of one of the HTY strains was used to localize the origin of the cloned telomeric DNA by in situ hybridization to the tip of the long arm of chromosome 7.
- OSTI ID:
- 6984024
- Journal Information:
- Proceedings of the National Academy of Sciences of the United States of America; (USA), Journal Name: Proceedings of the National Academy of Sciences of the United States of America; (USA) Vol. 86:16; ISSN 0027-8424; ISSN PNASA
- Country of Publication:
- United States
- Language:
- English
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59 BASIC BIOLOGICAL SCIENCES
ANIMALS
BIOLOGICAL MARKERS
BIOLOGICAL MATERIALS
BLOOD
BLOOD CELLS
BODY FLUIDS
CHROMOSOMES
CLONING
DNA HYBRIDIZATION
DNA-CLONING
EUMYCOTA
FUNGI
GENETIC MAPPING
HETEROCHROMOSOMES
HYBRIDIZATION
LEUKOCYTES
MAMMALS
MAN
MAPPING
MATERIALS
MICROORGANISMS
PLANTS
PRIMATES
SACCHAROMYCES
SACCHAROMYCES CEREVISIAE
VERTEBRATES
YEASTS