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In Vitro Translation of mRNA in a Rabbit Reticulocyte Lysate Cell-Free System

  • Louise Olliver
  • Charles D. Boyd
Protocol
  • 117 Downloads
Part of the Springer Protocols Handbooks book series (SPH)

Abstract

The identification of specific messenger RNA molecules and the characterization of the proteins encoded by them has been greatly assisted by the development of in vitro translation systems. These cell-free extracts comprise the cellular components necessary for protein synthesis, i.e., ribosomes, tRNA, rRNA, amino acids, initiation, elongation and termination factors, and the energy-generating system (1). Heterologous mRNAs are faithfully and efficiently translated in extracts of HeLa cells (2), Krebs II ascites tumor cells (2), mouse L cells (2), rat and mouse liver cells (3), Chinese hamster ovary (CHO) cells (2), and rabbit reticulocyte lysates (2,4), in addition to those of rye embryo (5) and wheat germ (6). Translation in cell-free systems is simpler and more rapid (60 min vs 24 h) than the in vivo translation system using Xenopus oocytes.

Keywords

Translation Product Translation System Microsomal Membrane Label Amino Acid Potassium Acetate 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Humana Press Inc., Totowa, NJ 2000

Authors and Affiliations

  • Louise Olliver
    • 1
  • Charles D. Boyd
    • 1
  1. 1.University of Medicine and DentistryNew Brunswick

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