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Nucleic Acids Research, 1985, Vol. 13, No. 10 3495-3514
© 1985

Articles

Aberrant recombination events in B cell lines derived from a x-deficient human

Janet Stavnezer, Olga Kekish, David Batter*, Joseph Grenier+, Ivan Balaz§, Earl Henderson1 and Ben J. M. Zegers2

Molecular Biology and Virology Program, Sloan-Kettering Institute New York, NY 10021 1Department of Microbiology and Immunology, Temple University Philadelphia, PA 19140, USA 2Department of Immunology, University Children's Hospital Nieuwe Gracht 137, Utrecht 66312, The Netherlands

Received February 13, 1985. Revised April 11, 1985. Accepted April 16, 1985.

We have analyzed the structure of Ig {kappa} chain genes in B cell lines derived from a hiinan individual who cannot synthesize any {kappa} chains, and whose Igs all contain {lambda} chains (1). We have characterized secondary DNA recombination events at two {kappa} alleles which have undergone misaligned V-J recombinations. One such secondary recombination has joined the flanking sequences of a V{kappa} and a J{kappa}2 gene segment as if it were the reciprocal product of a V-J{kappa}2 recombination, and resulted in the displacement of the recombined VJ{kappa}1 gene segments from the C{kappa} locus. The non-rearranged form of the V{kappa} fragment which had recombined with the J{kappa}2 flank was cloned. Nucleotide sequencing of this fragment identified a V{kappa} gene that differed by at least 38% from all previously sequenced human V{kappa} genes. The other V-J{kappa} segment analyzed has undergone a secondary recombination at a different site from that described above, at a site within the intervening sequence between the J{kappa} and C{kappa} gene segments, similar to the location of secondary recombinations which have occurred in {lambda} + B cell lines from mice and humans (2, 3). These results prove that multiple recoiiibinations can occur at one J{kappa}-C{kappa} locus.

*Present address: Western Psychiatric Institute of Pittsburgh, Pittsburgh, PA 15213, USA

+Present address: McGill University Medical School, Montreal, Canada

§Present address: Actagen Incorp., 4 Westchester Plaza, Elmsford, NY 10523, USA


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