Integrative selection of human chromosome-specific yeast artificial chromosomes

Journal Article · Sun Sep 01 04:00:00 EDT 1991 · Proceedings of the National Academy of Sciences of the United States of America; (United States)

DOI:https://doi.org/10.1073/pnas.88.17.7788· OSTI ID:5703021

Pavan, W J; Reeves, R G ^[1]

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1. Johns Hopkins Univ., Baltimore, MD (United States)

Human specific integrative selection vectors (ISVs) were designed to optimize integration of a yeast-selectable marker specifically into yeast artificial chromosomes (YACs) derived from human but not mouse DNA. ISVs were transformed into a YAC genomic library constructed from DNA of a human-mouse somatic cell hybrid containing chromosome 21 (HSA21) as the only human chromosome. One percent of the yeast in the original library contained HSA21-derived YACs; between 45% and 54% of the yeast recovered after transformation with ISV vectors contained human YACs. Integrative selection provides a rapid means of obtaining a highly enriched population of human chromosome-specific YACs by eliminating the labor-intensive steps of isolating and screening primary transformants. The procedure is biased toward the selection of YACs that contain a large number of targets for homologous recombination; thus, libraries constructed by this procedure will be composed primarily of the largest YACs in the population.

• OSTI ID: 5703021
• Journal Information: Proceedings of the National Academy of Sciences of the United States of America; (United States), Journal Name: Proceedings of the National Academy of Sciences of the United States of America; (United States) Vol. 88:17; ISSN 0027-8424; ISSN PNASA
• Country of Publication: United States
• Language: English

Related Subjects

550200* -- Biochemistry
59 BASIC BIOLOGICAL SCIENCES
ANIMALS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
CHROMOSOMES
CLONING
CYTOLOGICAL TECHNIQUES
DAYS LIVING RADIOISOTOPES
DNA HYBRIDIZATION
DNA-CLONING
ELECTROPHORESIS
EUMYCOTA
FUNGI
HYBRIDIZATION
ISOTOPES
LIGHT NUCLEI
MAMMALS
MAN
MICROORGANISMS
NUCLEI
ODD-ODD NUCLEI
PHOSPHORUS 32
PHOSPHORUS ISOTOPES
PLANTS
PRIMATES
RADIOISOTOPES
VERTEBRATES
YEASTS