Proceedings of the National Academy of Sciences · 1983 · 399 citations · 26 references
Life SciencesCell BiologyMolecular BiologyGene ExpressionGenetic EngineeringProtein EngineeringAntibody Expression SystemsEnzyme ExpressionHybridoma TechnologyImmunoglobulin Gene RearrangementProtein Expression EngineeringProtein SynthesisHybridoma Cell LinesImmunoglobulin Gene ExpressionImmunoglobulin Light Chain RearrangementImmunoglobulin ProductionMyeloma Cell LinesPlasmid Vector ConstructionTransgene ExpressionTwo-dimensional Gel Electrophoresis
Myeloma, hybridoma, and thymoma cell lines have been successfully transfected for the Escherichia coli xanthine-guanine phosphoribosyltransferase gene (gpt) by using the plasmid vector pSV2-gpt. The transformed cells synthesize the bacterial enzyme 5-phospho-alpha-D-ribose-1-diphosphate:xanthine phosphoribosyltransferase (XGPRT; EC 2.4.2.22) and have been maintained in selective medium for over 4 months. Lymphoid cell lines expressing a K immunoglobulin light chain were obtained by transfecting cells with pSV2-gpt containing a rearranged K light chain genomic segment from the S107 myeloma cell line. The S107 light chain is synthesized in gpt-transformed J558L myeloma cells and is identical to the light chain synthesized by the S107 myeloma cell line, as judged by immunoprecipitation and two-dimensional gel electrophoresis. Furthermore, this light chain is synthesized and secreted as part of an intact antibody molecule by transformed hybridoma cells that normally secrete an IgGl (gamma, K) antibody molecule. No light chain synthesis was detected in a similarly transformed rat myeloma or a mouse thymoma line.
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Properties of Monoclonal Antibodies to Mouse Ig Allotypes, H-2, and Ia Antigens
Vernon T. Oi, Pauline Jones, James W. Goding et al. · 1979 · 979 citations
Expression of a Bacterial Gene in Mammalian Cells
Richard Mulligan, Paul Berg · Science · 1980 · 785 citations