Extrachromosomal DNA substrates in pre-B cells undergo inversion or deletion at immunoglobulin V-(D)-J joining signals.
Sequences encoding immunoglobulin variable domains are known to be assembled from variable (V), diversity (D), and joining (J) segments by site-specific recombination. We present a sensitive and rapid assay for V-(D)-J recombination that uses plasmid DNA transiently introduced into transformed pre-B cells, and demonstrates that the recombination is independent of any unique chromosomal context. Sequences sufficient to constitute recombination sites are contained within the 84 and 42 bp flanking, respectively, the murine J kappa 1 and V kappa L8 segments, which include the known heptamer-nonamer V-(D)-J joining signals. Deletion and inversion occur at comparable frequencies. Thus, V-(D)-J recombination may be relatively insensitive to the topological arrangement of sites, and events at the two novel junctions produced by the reaction may be coupled.
D000818:Animals; D001402:B-Lymphocytes / Q000502:physiology*; D002478:Cells, Cultured; D002872:Chromosome Deletion; D007446:Chromosome Inversion; D007135:Immunoglobulin Variable Region / Q000235:genetics*; D051379:Mice; D010957:Plasmids; D011995:Recombination, Genetic; D013329:Structure-Activity Relationship
D007135:Immunoglobulin Variable Region