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

Biochemistry. 1976 Jun 29;15(13):2775-9. doi: 10.1021/bi00658a011.

Abstract

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.

MeSH terms

  • Cell Line
  • Cell-Free System
  • Hot Temperature
  • Immunoglobulin Light Chains / biosynthesis*
  • Immunoglobulin lambda-Chains / biosynthesis*
  • Kinetics
  • Multiple Myeloma / metabolism*
  • Nucleic Acid Denaturation
  • Nucleic Acid Hybridization
  • Polyribosomes / metabolism
  • Protein Biosynthesis*
  • RNA, Messenger / isolation & purification
  • RNA, Messenger / metabolism*
  • RNA-Directed DNA Polymerase / metabolism

Substances

  • Immunoglobulin Light Chains
  • Immunoglobulin lambda-Chains
  • RNA, Messenger
  • RNA-Directed DNA Polymerase