In vitro analysis of translation enhancers

Methods Mol Biol. 2008:451:113-24. doi: 10.1007/978-1-59745-102-4_8.

Abstract

The genomes of many plant viruses contain translation-enhancing sequences that allow them to compete successfully with host messenger RNAs for the translation machinery. Identification of translation enhancer elements is valuable, both to gain understanding of virus gene expression control and to apply them as tools for engineering gene expression in plant cells. Here, we describe experiments designed to detect viral elements that enhance translation, focusing on cap-independent translation activity, using a high fidelity cell-free wheat germ translation extract.

MeSH terms

  • Enhancer Elements, Genetic / genetics*
  • Genes, Reporter
  • Genetic Techniques*
  • Genome, Viral*
  • Open Reading Frames
  • Plant Viruses / genetics*
  • Protein Biosynthesis*
  • RNA, Messenger / genetics*
  • RNA, Viral / genetics
  • Seeds
  • Triticum / genetics

Substances

  • RNA, Messenger
  • RNA, Viral