Reciprocal homologous recombination in or near antibody VDJ genes

Asad Umar, Patricia J. Gearhart

Research output: Contribution to journalArticle

Abstract

To determine if rearranged heavy chain variable (VDJ) genes can recombine with each other by crossing over of DNA strands, we constructed a transgene that contained a promoter, VDJ gene, reporter gene to detect crossover events, intron enhancer, matrix attachment region, and constant gene for IgM (Cμ). Following immunization of transgenic mice, hybrid molecules were isolated from B cell DNA which contained the transgene recombined with the endogenous IgH locus. Reciprocal products of crossovers were detected by plasmid rescue and PCR amplification, and they were sequenced. Recombination occurred somewhere within 147 bp of homology that contained the J(H)4 gene segment and 3' flanking DNA. The recombined transgenes had a 20-fold increase in mutation in the VDJ region compared to nonrecombined transgenes, which indicates that DNA sequences 3' of the Cμ gene in the endogenous IgH locus are necessary for full activity of the mutator mechanism. The recovery of recombinants between VDJ transgenes and endogenous VDJ genes raises the possibility that reciprocal recombination may somatically diversify rearranged genes between maternal and paternal alleles.

Original languageEnglish (US)
Pages (from-to)2392-2400
Number of pages9
JournalEuropean Journal of Immunology
Volume25
Issue number8
DOIs
StatePublished - Aug 1995

Fingerprint

Homologous Recombination
Transgenes
Antibodies
Genes
Genetic Recombination
DNA
Matrix Attachment Regions
Reporter Genes
Introns
Transgenic Mice
Immunoglobulin M
Immunization
Plasmids
B-Lymphocytes
Alleles
Mothers
Polymerase Chain Reaction
Mutation

Keywords

  • Homologous recombination
  • IgH locus
  • Immunoglobulin transgenes
  • Somatic mutation

ASJC Scopus subject areas

  • Immunology

Cite this

Reciprocal homologous recombination in or near antibody VDJ genes. / Umar, Asad; Gearhart, Patricia J.

In: European Journal of Immunology, Vol. 25, No. 8, 08.1995, p. 2392-2400.

Research output: Contribution to journalArticle

Umar, Asad ; Gearhart, Patricia J. / Reciprocal homologous recombination in or near antibody VDJ genes. In: European Journal of Immunology. 1995 ; Vol. 25, No. 8. pp. 2392-2400.
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