Generation of deletion derivatives by targeted transformation of human-derived yeast artificial chromosomes

Journal Article · Thu Feb 01 04:00:00 EST 1990 · Proceedings of the National Academy of Sciences of the United States of America; (USA)

DOI:https://doi.org/10.1073/pnas.87.4.1300· OSTI ID:6275375

Pavan, W J; Hieter, P; Reeves, R H ^[1]

---

1. Johns Hopkins Univ. School of Medicine, Baltimore, MD (USA)

Mammalian DNA segments cloned as yeast artificial chromosomes (YACs) can be manipulated by DNA-mediated transformation when placed in an appropriate yeast genetic background. A fragmenting vector has been developed that can introduce a yeast telomere and selectable marker into human-derived YACs at specific sites by means of homologous recombination, deleting all sequences distal to the recombination site. A powerful application of the method uses a human Alu family repeat sequence to target recombination to multiple independent sites on a human-derived YAC. Sets of deletion derivatives generated by this procedure greatly facilitate restriction mapping of large genomic segments. Targeting recombination with single copy sequences, such as cDNAs, will have many additional applications. This approach establishes a paradigm for manipulation and characterization of mammalian DNA segments cloned as YACs.

• OSTI ID: 6275375
• 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. 87:4; ISSN 0027-8424; ISSN PNASA
• Country of Publication: United States
• Language: English

Related Subjects

550400* -- Genetics
59 BASIC BIOLOGICAL SCIENCES
ANIMALS
CELL CONSTITUENTS
CHROMOSOMAL ABERRATIONS
CHROMOSOMES
CLONING
DNA
DNA HYBRIDIZATION
DNA SEQUENCING
DNA-CLONING
ELECTROPHORESIS
EUMYCOTA
FUNGI
GENE RECOMBINATION
HETEROCHROMOSOMES
HYBRIDIZATION
MAMMALS
MAN
MICROORGANISMS
MUTATIONS
NUCLEIC ACIDS
ORGANIC COMPOUNDS
PLANTS
PLASMIDS
PRIMATES
RECOMBINANT DNA
SACCHAROMYCES
SACCHAROMYCES CEREVISIAE
STRUCTURAL CHEMICAL ANALYSIS
VERTEBRATES
YEASTS