Analysis of large deletions in the HPRT gene of primary human fibroblasts using the polymerase chain reaction

Journal Article · Fri Jan 01 04:00:00 EST 1993 · Somatic Cell and Molecular Genetics; (United States)

DOI:https://doi.org/10.1007/BF01233950· OSTI ID:6616225

Morris, T; Masson, W; Singleton, B; Thacker, J ^[1]

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1. MRC Radiobiology Unit, Chilton (United Kingdom)

Spontaneous and X-ray-induced mutants of the HPRT gene were isolated from two primary human fibroblast lines. The limited life-span of the mutants restricted the use of methods requiring large quantities of DNA, and the polymerase chain reaction (PCR) was used in particular to check for the presence of multiple genomic sites in mutant analysis. Robust PCR primers were designed to amplify sites of up to 1 kb, mostly with 1-kb spacings between sites, over the entire 56-kb HPRT gene region. Using PCR, large deletions were found in 43% of independent X-ray-induced mutants, and their breakpoints were localized where these fell within the gene. Anonymous DNA sites in the Xq26 chromosomal region containing HPRT (covering [ge]1.5 Mb) were also amplified by PCR to assess codeletion with HPRT; sites up to 1 Mb distal to the gene (DXS86, DXS10) were codeleted in some mutants, but no mutant was found with loss of a proximal site (DXS79). 51 refs., 5 figs., 2 tabs.

• OSTI ID: 6616225
• Journal Information: Somatic Cell and Molecular Genetics; (United States), Journal Name: Somatic Cell and Molecular Genetics; (United States) Vol. 19:1; ISSN 0740-7750; ISSN SCMGDN
• Country of Publication: United States
• Language: English

Related Subjects

550400* -- Genetics
59 BASIC BIOLOGICAL SCIENCES
AMPLIFICATION
BIOLOGICAL EFFECTS
CHROMOSOMES
DNA
DNA SEQUENCING
ELECTROMAGNETIC RADIATION
ENZYMES
GENE MUTATIONS
GENETIC MAPPING
HETEROCHROMOSOMES
HUMAN CHROMOSOMES
HUMAN X CHROMOSOME
IONIZING RADIATIONS
MAPPING
MUTATIONS
NUCLEIC ACIDS
ORGANIC COMPOUNDS
PROTEINS
RADIATIONS
STRUCTURAL CHEMICAL ANALYSIS
TRANSFERASES
X CHROMOSOME
X RADIATION