Development of a thermostable β-glucuronidase-based reporter system for monitoring gene expression in hyperthermophiles

Biotechnol Bioeng. 2012 Jul;109(7):1881-6. doi: 10.1002/bit.24432. Epub 2012 Jan 11.

Abstract

Mesophilic glucuronidases are the most widely used reporters of gene expression in plants, but unsuitable as reporters in (hyper-)thermophiles due their insufficient thermal stability. Here we present the native 66.8 kDa thermostable β-glucuronidase of Sulfolobus solfataricus. The enzyme activity is characterized in a wide temperature range ideal for, but not limited to, in vivo genetic study of hyperthermophiles. As a proof of concept, we demonstrate its use as a reporter of gene expression in Sulfolobus, by monitoring a promoter fusion created with the β-glucuronidase coding gene gusB and a copper-responsive promoter.

Publication types

  • Letter
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cloning, Molecular
  • Gene Expression Profiling*
  • Genes, Reporter*
  • Glucuronidase / genetics*
  • Glucuronidase / metabolism
  • Promoter Regions, Genetic
  • Protein Stability
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Sulfolobus solfataricus / enzymology*
  • Sulfolobus solfataricus / genetics
  • Temperature

Substances

  • Recombinant Proteins
  • Glucuronidase