Purification of immunuglobulin light chain messenger RNA by immunoprecipitation from mouse myeloma tumor, MOPC-31C.
Polysomes producing IgGl(kappa) myeloma protein were specifically selected by an immunoprecipitation method, and immunoglobulin light chain mRNA was purified from the precipitated polysomes. The purified mRNA migrated predominantly as a single band and the molecular weight of this mRNA was calculated to be 410.000 by polyacrylamide gel electrophoresis in 98% formamide. A protein possessing a molecular weight of 25,000, which is the size of the light chain precursor, was synthesized as a major product of translation in a wheat germ cell-free system. DNA complementary to the mRNA (cDNA) was prepared with avian myeloblastosis virus RNA-dependent DNA polymerase. This cDNA had an average size of 8.3S as determined by sedimentation through an alkaline sucrose gradient. Using this cDNA, Crt 1/2 values of template RNA and RNA from various preparations were calculated from the results of molecular hybridization. The relative content of the mRNA increased 4,4-fold during the immunoprecipitation of polysomes.
D000818:Animals; D009189:Avian Myeloblastosis Virus / Q000201:enzymology; D002478:Cells, Cultured; D004247:DNA / Q000096:biosynthesis; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism; D007147:Immunoglobulin Light Chains / Q000096:biosynthesis*; D007145:Immunoglobulin kappa-Chains / Q000096:biosynthesis*; D008715:Methionine / Q000378:metabolism; D051379:Mice; D008970:Molecular Weight; D009101:Multiple Myeloma / Q000032:analysis*; D009194:Myeloma Proteins / Q000096:biosynthesis; D009693:Nucleic Acid Hybridization; D011132:Polyribosomes; D012333:RNA, Messenger / Q000302:isolation & purification* / Q000378:metabolism
D007147:Immunoglobulin Light Chains; D007145:Immunoglobulin kappa-Chains; D009194:Myeloma Proteins; D012333:RNA, Messenger; D004247:DNA; D008715:Methionine; D004259:DNA-Directed DNA Polymerase