Somatic Recombination of Heavy Chain Variable Region Transgenes with the Endogenous Immunoglobulin Heavy Chain Locus in Mice

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 89; no. 21; pp. 10321 - 10325
• Main Authors: Giusti, Angela M., Coffee, Richard, Manser, Tim
• Format: Journal Article
• Language: English
• Published: Washington, DC National Academy of Sciences of the United States of America 01.11.1992; National Acad Sciences; National Academy of Sciences
• Subjects: Animals; Antibodies; Antibodies - blood; Antibodies - genetics; B lymphocytes; Base Sequence; Biological and medical sciences; Biotechnology; Blotting, Southern; Cloning, Molecular; Deoxyribonucleic acid; DNA; DNA - genetics; DNA - isolation & purification; Female; Fundamental and applied biological sciences. Psychology; Genes, Immunoglobulin; Genetic engineering; Genetic loci; Genetic technics; Genetic vectors; Hybridity; Hybridomas; Immunity (Disease); Immunoglobulin Heavy Chains - genetics; Immunoglobulin Variable Region - genetics; Male; Medical research; Methods. Procedures. Technologies; Mice; Mice, Inbred A; Mice, Transgenic; Molecular Sequence Data; Restriction Mapping; Rodents; Transgenes; Transgenic animals; Transgenic animals and transgenic plants; Immunoglobulins; Recombination; Antibody; heavy-Peptide chain; Variable region; Rodentia; Transgenic animal; Hybridoma; B-Lymphocyte; Immunogenetics; Vertebrata; Mammalia; Gene; Mouse; Constant region
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.89.21.10321

Transgenic lines of mice were derived by using plasmid constructs containing DNA encoding an antibody heavy chain variable-diversity-joining region (VH-D-JH) and various amounts of 5' and 3' flanking DNA but lacking any repetitive isotype switch (S) or constant (C) region DNA. Unexpectedly, many of the antibody VHregions expressed by B-cell hybridomas generated from immunized transgenic mice were found to be of transgenic origin. Further analyses showed that somatic events had generated hybrid genomic loci in the mice containing the transgenic VH-D-JHgene and plasmid sequences 5' of endogenous heavy chain C region genes. Thus, VH-D-JHtransgenes lacking S and C region DNA can recombine with endogenous Igh DNA, leading to the expression of transgene-encoded antibody.