Construction of mouse chromosome-specific DNA libraries and their use for the detection of X-ray-induced aberrations.
We describe here the development of mouse chromosome-specific DNA libraries and their use in the detection of radiation-induced chromosome aberrations by fluorescence in situ hybridization. Large metacentric chromosomes, resulting from a translocation involving chromosomes 1, 11 and 13, were flow-sorted. Using a slit-scan technique for morphometric analysis, metacentric chromosomes were separated from normal acrocentric chromosomes and their aggregates. DNA from the metacentric chromosomes was amplified by PCR using the linker/adaptor method. In this pilot study, mouse was whole-body irradiated with 1, 2 and 3 Gy and aberrations were scored in metaphase spreads of splenocytes cultured in vitro. The results indicate that directly after radiation exposure, stable and unstable aberrations are induced at about equal frequencies in the splenocytes. The availability of chromosome-specific probes for mouse may prove very useful when analysing the behaviour of stable aberrations, as well as the testing of many suspected mutagenic carcinogens and aneugens in vivo for induction of chromosomal translocations and non-disjunction, respectively.
Boei
J J
JJ
Department of Radiation Genetics and Chemical Mutagenesis, State University of Leiden, The Netherlands.
Balajee
A S
AS
de Boer
P
P
Rens
W
W
Aten
J A
JA
Mullenders
L H
LH
Natarajan
A T
AT
D000818:Animals; D001483:Base Sequence; D002869:Chromosome Aberrations; D004247:DNA / Q000235:genetics* / Q000528:radiation effects*; D015342:DNA Probes; D005260:Female; D015698:Genomic Library; D017403:In Situ Hybridization; D007621:Karyotyping; D008677:Metaphase; D051379:Mice; D008969:Molecular Sequence Data; D010865:Pilot Projects; D016133:Polymerase Chain Reaction; D013154:Spleen / Q000166:cytology / Q000528:radiation effects; D014178:Translocation, Genetic; D014916:Whole-Body Irradiation
D015342:DNA Probes; D004247:DNA