New unstable variants of green fluorescent protein for studies of transient gene expression in bacteria

Appl Environ Microbiol. 1998 Jun;64(6):2240-6. doi: 10.1128/AEM.64.6.2240-2246.1998.

Abstract

Use of the green fluorescent protein (Gfp) from the jellyfish Aequorea victoria is a powerful method for nondestructive in situ monitoring, since expression of green fluorescence does not require any substrate addition. To expand the use of Gfp as a reporter protein, new variants have been constructed by the addition of short peptide sequences to the C-terminal end of intact Gfp. This rendered the Gfp susceptible to the action of indigenous housekeeping proteases, resulting in protein variants with half-lives ranging from 40 min to a few hours when synthesized in Escherichia coli and Pseudomonas putida. The new Gfp variants should be useful for in situ studies of temporal gene expression.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • DNA Primers / genetics
  • Escherichia coli / genetics*
  • Gene Expression
  • Genes, Reporter
  • Genetic Variation*
  • Green Fluorescent Proteins
  • Half-Life
  • Luminescent Proteins / genetics*
  • Luminescent Proteins / metabolism
  • Molecular Sequence Data
  • Pseudomonas putida / genetics*
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Scyphozoa / genetics

Substances

  • DNA Primers
  • Luminescent Proteins
  • Recombinant Proteins
  • Green Fluorescent Proteins