In vitro translation in a hybrid cell free lysate with exogenous cellular ribosomes

Biochem J. 2015 May 1;467(3):387-98. doi: 10.1042/BJ20141498.

Abstract

Cell free protein synthesis systems (CFPS) have been widely used to express proteins and to explore the pathways of gene expression. In the present manuscript, we describe the design of a novel adaptable hybrid in vitro translation system which is assembled with ribosomes isolated from many different origins. We first show that this hybrid system exhibits all important features such as efficiency, sensitivity, reproducibility and the ability to translate specialized mRNAs in less than 1 h. In addition, the unique design of this cell free assay makes it highly adaptable to utilize ribosomes isolated from many different organs, tissues or cell types.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 5' Untranslated Regions
  • Animals
  • Cell Line
  • Cell-Free System
  • Cells, Cultured
  • Cricetinae
  • HeLa Cells
  • Humans
  • In Vitro Techniques
  • Jurkat Cells
  • Luciferases, Renilla / biosynthesis
  • Luciferases, Renilla / genetics
  • Mice
  • Poliovirus / genetics
  • Protein Biosynthesis* / genetics
  • RNA, Messenger / genetics
  • Rabbits
  • Reticulocytes / metabolism
  • Ribosomes / metabolism*
  • beta-Globins / biosynthesis
  • beta-Globins / genetics

Substances

  • 5' Untranslated Regions
  • RNA, Messenger
  • beta-Globins
  • Luciferases, Renilla