Cloning human telomeric DNA fragments into Saccharomyces cerevisiae using a yeast-artificial-chromosome vector

Journal Article · Tue Aug 01 00:00:00 EDT 1989 · Proceedings of the National Academy of Sciences of the United States of America; (USA)

DOI:https://doi.org/10.1073/pnas.86.16.6240· OSTI ID:6984024

Riethman, H C; Burke, D T; Olson, M V ^[1]; Moyzis, R K; Meyne, J ^[2]

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1. Washington Univ. School of Medicine, St. Louis, MO (USA)
2. 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), Vol. 86:16; ISSN 0027-8424
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
HETEROCHROMOSOMES
DNA HYBRIDIZATION
SACCHAROMYCES CEREVISIAE
DNA-CLONING
BIOLOGICAL MARKERS
GENETIC MAPPING
LEUKOCYTES
MAN
ANIMALS
BIOLOGICAL MATERIALS
BLOOD
BLOOD CELLS
BODY FLUIDS
CHROMOSOMES
CLONING
EUMYCOTA
FUNGI
HYBRIDIZATION
MAMMALS
MAPPING
MATERIALS
MICROORGANISMS
PLANTS
PRIMATES
SACCHAROMYCES
VERTEBRATES
YEASTS
550200* - Biochemistry