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Interstrain conservation of the murine GAT‐specific antibody V kappa repertoire as analyzed at the germline gene level.
Author(s) -
Schiff C.,
Corbet S.,
Milili M.,
Fougereau M.
Publication year - 1983
Publication title -
the embo journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.484
H-Index - 392
eISSN - 1460-2075
pISSN - 0261-4189
DOI - 10.1002/j.1460-2075.1983.tb01656.x
Subject(s) - complementary dna , biology , kappa , microbiology and biotechnology , genetics , germline , gene , immunoglobulin light chain , cdna library , antibody , stop codon , mathematics , geometry
A cDNA library was constructed in pBR322 from mRNA encoding an anti‐GAT (Glu60 Ala30 Tyr10) monoclonal antibody kappa chain. Two cDNA clones were extensively characterized. One, L XI 62, was derived from an aberrant V kappa‐J kappa rearrangement which resulted in a frame‐shift at position 96, leading to a stop codon at the very beginning of the constant region. The second, L XIX 27, 1150 bp long, was unequivocally assigned to a GAT‐specific kappa chain, by comparison of its nucleotide sequence with the previously determined NH2‐terminal amino acid sequence of the isolated kappa chain. A specific probe, containing the leader and most of the V kappa gene‐encoded region, was prepared from this clone and hybridized to EcoRI and BamHI restriction fragments of liver (unrearranged) DNA extracted from the BALB/c, DBA/2 and C57BL/6 mouse strains. Under stringent conditions, similar patterns were observed for all three strains, and consisted of a small number of bands (3‐5). Under nonstringent conditions, patterns were again very similar when the different strains were compared, although 15‐20 bands could be identified. These observations support the hypothesis that the GAT‐specific kappa chains found in antibodies expressing the public CGAT idiotypes are encoded by a very small number of germline genes. This V kappa repertoire seems extremely conserved between the three strains that were analyzed, an observation which correlates with the interstrain conservation of these public idiotypic specificities.