DNA-mediated transfer of human melanoma cell surface glycoprotein gp130: identification of transfectants by erythrocyte rosetting.
DNA sequences encoding a human melanoma membrane-bound sialoglycoprotein of 130,000 molecular weight (gp130) were introduced into a clonal derivative of mouse B-16 melanoma cells with the selectable neomycin resistance gene (aminoglycoside phosphotransferase). Mouse transfectants were identified by a rapid and precise screening method with mouse monoclonal antibodies and erythrocyte rosetting. The frequency of gp130 transfectants was approximately 1 in 2,000 to 5,000 colonies with neo+ cells. Analysis of secondary mouse transfectants has revealed that the transfected gp130 has a molecular weight, isoelectric point, intracellular processing, peptide map, and spatial orientation of surface-exposed epitopes indistinguishable from those seen with gp130 from human melanoma cells. In contrast to primary transfectants, secondary transfectants expressing gp130 lack demonstrable human repetitive sequences.
D000818:Animals; D002478:Cells, Cultured; D004273:DNA, Neoplasm / Q000235:genetics*; D005786:Gene Expression Regulation; D006023:Glycoproteins / Q000235:genetics*; D006801:Humans; D008545:Melanoma / Q000235:genetics*; D051379:Mice; D008970:Molecular Weight; D009363:Neoplasm Proteins / Q000235:genetics*; D011499:Protein Processing, Post-Translational; D012397:Rosette Formation; D014162:Transfection
D004273:DNA, Neoplasm; D006023:Glycoproteins; D009363:Neoplasm Proteins