The recombination mediated by double-strand breaks in extrachromosomal DNA substrate carrying mouse immunoglobulin switch regions S mu and S gamma 2b

Abstract

Recombination in mouse cells was analyzed using extrachromosomal DNA substrates carrying the mouse immunoglobulin switch regions S mu and S gamma 2b. Recombination was detected at a frequency of 10(-2)-10(-3) in mouse fibroblasts and in pre-B cell lines, but at a low frequency in a scid fibroblast cell line. Restriction enzyme digestion profile revealed that most recombination occurred between the CMV promoter region, which neighbors the S mu upstream region, and the S gamma 2b region. However, frequency of direct recombination between the CMV promoter region and the S gamma 2b region was low as measured by the substrate-lacking S mu region. Nucleotide sequence analysis showed that recombination occurred between several homologous base-pairs, and extranucleotides were frequently found at the recombination junctions. These results indicate that recombination took the form of the recombination mediated by double-strand breaks. Double-strand breaks likely occurred in the S mu and/or S gamma 2b region, and the ends joined.