In vitro activation of a nonproductive immunoglobulin allele by a single base pair insertion

We have shown that one of the alleles in a hybridoma was nonproductive because the sequence in the N region of the heavy chain caused it to be out of frame and to terminate prematurely. This allele became productive, and the gamma 1 heavy chain that it encoded was secreted when an adenosine was inse...

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Bibliographic Details
Published inHybridoma Vol. 13; no. 4; p. 257
Main Authors Kobrin, B J, Schiff, C, Zivion, D, Scharff, M D, Spira, G
Format Journal Article
LanguageEnglish
Published United States 01.08.1994
Subjects
Alleles
Amino Acid Sequence
Base Sequence
Cells, Cultured
Cloning, Molecular
Enzyme-Linked Immunosorbent Assay
Gene Rearrangement, B-Lymphocyte - genetics
Hybridomas - immunology
Immunoglobulin G - genetics
Immunoglobulin Heavy Chains - genetics
Immunoglobulin kappa-Chains - genetics
Immunoglobulin Variable Region - genetics
Molecular Sequence Data
Mutagenesis, Insertional - genetics
Mutagenesis, Site-Directed - genetics
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Summary:We have shown that one of the alleles in a hybridoma was nonproductive because the sequence in the N region of the heavy chain caused it to be out of frame and to terminate prematurely. This allele became productive, and the gamma 1 heavy chain that it encoded was secreted when an adenosine was inserted to produce an open reading frame. This event occurred at a very low frequency following mutagenesis of the cultured cells. This result suggests that similar sorts of events must occur in vivo when both productive and nonproductive alleles undergo frequent mutations and that new genes may be expressed in hybridomas as they are being subcloned.
ISSN:0272-457X
DOI:10.1089/hyb.1994.13.257