Purification and translation of an immunoglobulin λ chain messenger RNA from mouse myeloma

Abstract
The 500-fold purification of an mRNA encoding an immunoglobulin .lambda. L chain derived from the mouse myeloma tumor, RPC-20 is described. 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 5 or 6 amino acids longer than the mature form of RPC-20 L 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 L chain.

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