Construction of human chromosome 21-specific yeast artificial chromosomes

Journal Article · Fri Dec 01 00:00:00 EST 1989 · Proceedings of the National Academy of Sciences of the United States of America; (USA)

DOI:https://doi.org/10.1073/pnas.86.24.9991· OSTI ID:6855056

McCormick, M K; Shero, J H; Hieter, P A; Antonarakis, S E ^[1]; Cheung, Meichi; Kan, Yuetwai ^[2]

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1. Johns Hopkins Univ. School of Medicine, Baltimore, MD (USA)
2. Univ. of California, San Francisco (USA)

Chromosome 21-specific yeast artificial chromosomes (YACs) have been constructed by a method that performs all steps in agarose, allowing size selection by pulsed-field gel electrophoresis and the use of nanogram to microgram quantities of DNA. The DNA sources used were hybrid cell line WAV-17, containing chromosome 21 as the only human chromosome and flow-sorted chromosome 21. The transformation efficiency of ligation products was similar to that obtained in aqueous transformations and yielded YACs with sizes ranging from 100 kilobases (kb) to > 1 megabase when polyamines were included in the transformation procedure. Twenty-five YACs containing human DNA have been obtained from a mouse-human hybrid, ranging in size from 200 to > 1000 kb, with an average size of 410 kb. Ten of these YACs were localized to subregions of chromosome 21 by hybridization of RNA probes to a panel of somatic cell hybrid DNA. Twenty-one human YACs, ranging in size from 100 to 500 kb, with an average size of 150 kb, were obtained from {approx} 50 ng of flow-sorted chromosome 21 DNA. Three were localized to subregions of chromosome 21. YACs will aid the construction of a physical map of human chromosome 21 and the study of disorders associated with chromosome 21 such as Alzheimer disease and Down syndrome.

• OSTI ID: 6855056
• Journal Information: Proceedings of the National Academy of Sciences of the United States of America; (USA), Vol. 86:24; ISSN 0027-8424
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
HEREDITARY DISEASES
GENETIC MAPPING
HUMAN CHROMOSOME 21
DNA HYBRIDIZATION
CELL FLOW SYSTEMS
DNA
DOWNS SYNDROME
ELECTROPHORESIS
ESCHERICHIA COLI
MENTAL DISORDERS
MESSENGER-RNA
NERVOUS SYSTEM DISEASES
TRANSCRIPTION
YEASTS
BACTERIA
CHROMOSOMES
CONGENITAL MALFORMATIONS
DISEASES
EUMYCOTA
FUNGI
HYBRIDIZATION
MALFORMATIONS
MAPPING
MICROORGANISMS
NUCLEIC ACIDS
ORGANIC COMPOUNDS
PATHOLOGICAL CHANGES
PLANTS
RNA
550400* - Genetics