International Mammalian Genome Society

The 15th International Mouse Genome Conference (2001)


POSTER 104 - MAPPING, TIMING AND MECHANISM OF MEIOTIC RECOMBINATION IN MICE

Dr Bernard de Massy
CNRS
Institut de Genetique Humaine
UPR1142/CNRS
141 rue de la Cardonille
Montpellier
34396
France

Co-Author:  Guillon H
Institution: CNRS, Institut de Genetique Humaine

Mapping, timing and mechanism of meiotic recombination in mice

The mechanism of meiotic recombination is poorly described in mice, partly due to the absence of a direct assay to monitor recombination events. We have undertaken the development of a new and unique molecular assay for meiotic recombination. We have designed this assay in a previously defined meiotic recombination hot-spot, the Lmp2 hot-spot (Shiroishi et al., Immunogenetics, 1990). This hot-spot is haplotype specific, has a frequency of 1-2% as determined genetically (1000 fold higher than genome average) and is localized  within 1.5Kb at the 3' end of the Lmp2 gene. We have first identified all polymorphic sites in a large region around the hot-spot in various strains, used this information to design allele-specific oligonucleotides and used these oligos to set-up a PCR assay for the detection of recombinant molecules (cross-over products). This allowed us to measure the frequency and amplify independent recombinant molecules in germ cells. Mapping and analysis of recombinant molecules locates the highest density of exchanges within 200bp and strongly suggests that this hot-spot is an initiation site of meiotic recombination. In addition, our assay allowed us to perform the first temporal analysis of meiotic recombination in mice and to identify the meiotic stage at which cross-over products are generated.


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