Ribosome-Inactivation Display System

  • Satoshi Fujita
  • Jing-Min Zhou
  • Kazunari Taira
Part of the Methods in Molecular Biology™ book series (MIMB, volume 352)

Abstract

We present a novel strategy for the connection of phenotype and genotype in vitro that can be used for the selection of functional proteins. The strategy involves the generation of a stable complex among a ribosome, an messenger RNA and its translated protein, without removal of the termination codon, as a result of the action of the ricin A chain during translation. The technique requires no transfection, no chemical synthesis, no ligation, and no removal of the termination codon. Thus, our novel ribosome-inactivation display system should provide, without loss of the pool population, a reliable, simple, and robust selection system for the in vitro evolution of the properties of proteins in a predictable direction by a combination of randomization and appropriate selection strategies.

Key Words

Ribosome-inactivation display system in vitro selection molecular evolution protein ribosome display mRNA display phage display ricin A chain protein-ribosome-mRNA complex 

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Copyright information

© Humana Press Inc. 2007

Authors and Affiliations

  • Satoshi Fujita
    • 1
    • 2
  • Jing-Min Zhou
    • 1
    • 3
  • Kazunari Taira
    • 1
    • 3
  1. 1.Department of Chemistry and BiotechnologySchool of Engineering, The University of TokyoHongo, TokyoJapan
  2. 2.Research Institute for Cell EngineeringNational Institute of Advanced Industrial Science and Technology (AIST)TokyoJapan
  3. 3.Gene Function Research LaboratoryNational Institute of Advanced Industrial Science and Technology (AIST)Tsukuba Science CityJapan

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