Reassembling green fluorescent protein for in vitro evaluation of trans-translation

Nucleic Acids Res. 2020 Feb 28;48(4):e22. doi: 10.1093/nar/gkz1204.

Abstract

In order to discover new antibiotics with improved activity and selectivity, we created a reliable in vitro reporter system to detect trans-translation activity, the main mechanism for recycling ribosomes stalled on problematic messenger RNA (mRNA) in bacteria. This system is based on an engineered tmRNA variant that reassembles the green fluorescent protein (GFP) when trans-translation is active. Our system is adapted for high-throughput screening of chemical compounds by fluorescence.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anti-Bacterial Agents / chemical synthesis
  • Anti-Bacterial Agents / pharmacology
  • Bacteria / drug effects
  • Bacteria / genetics*
  • Green Fluorescent Proteins / genetics*
  • Humans
  • Protein Biosynthesis* / drug effects
  • RNA, Messenger / genetics*
  • RNA-Binding Proteins / genetics
  • Ribosomes / drug effects
  • Ribosomes / genetics

Substances

  • Anti-Bacterial Agents
  • RNA, Messenger
  • RNA-Binding Proteins
  • Green Fluorescent Proteins