Purification and translation of an immunoglobulin lambda chain messenger RNA from mouse myeloma.
Here we describe the 500-fold purification of an mRNA encoding an immunoglobulin lambda light chain derived from the mouse myeloma tumor, RPC-20. Purification involves the isolation of membrane-bound polysomes, oligo(dT)-cellulose chromatography, and sucrose gradient centrifugation under conditions favoring denaturation of polynucleotide complexes. The mRNA purified in this way directs the cell-free synthesis of a polypeptide which is five or six amino acids longer than the mature form of RPC-20 light chain. In addition to directing the synthesis of a precursor-like polypeptide, the mRNA migrates on electrophoresis as a band containing approximately 1150 nucleotides, about 500 more than required to encode the mature form of the light chain.
D002460:Cell Line; D002474:Cell-Free System; D006358:Hot Temperature; D007147:Immunoglobulin Light Chains / Q000096:biosynthesis*; D007146:Immunoglobulin lambda-Chains / Q000096:biosynthesis*; D007700:Kinetics; D009101:Multiple Myeloma / Q000378:metabolism*; D009691:Nucleic Acid Denaturation; D009693:Nucleic Acid Hybridization; D011132:Polyribosomes / Q000378:metabolism; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000302:isolation & purification / Q000378:metabolism*; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism
D007147:Immunoglobulin Light Chains; D007146:Immunoglobulin lambda-Chains; D012333:RNA, Messenger; D012194:RNA-Directed DNA Polymerase