Human Molecular Genetics, Vol 5, 875-886, Copyright © 1996 by Oxford University Press
ME Moynahan, E Akgun and M Jasin
Homologous recombination is a conserved process of genetic exchange
generated by homologous pairing of nucleotides. Species diversity and gene
evolution are dependent on the outcomes of recombination during germ cell
development, yet systems to study mammalian germline recombination,
especially those with applications to human genetics, are not well
developed. We report on a transgenic mouse system designed to study
recombination within test sequences in the male germline utilizing an
intron-interrupted lacZ reporter gene. beta-galactosidase positive sperm
are detected and quantitated by flow cytometry using fluorogenic
substrates. Examination of recombination within a 1.7 kb repeat of test
sequences derived from the human glycophorin breakpoint cluster region
detects approximately 0.04-0.09% fluorescent sperm. Confirmation that these
sperm result from recombination in the germline comes from histochemical
staining of testicular cells, examination of spliced mRNA, and PCR analysis
of sorted sperm populations. The system is readily adaptable to studies of
other sequences reported to have elevated levels of recombination,
including those implicated in human genetic disease. Investigations of the
molecular basis for genomic instability at specific chromosomal locations
may yield important insights into mechanisms of chromosomal loss and
rearrangements.
ARTICLES
A model for testing recombinogenic sequences in the mouse germline
Department of Medicine, Memorial Sloan-Kettering Cancer Center, New York, NY 10021, USA.
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