title: Development of a thermostable β-glucuronidase-based reporter system for monitoring gene expression in hyperthermophiles.
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.
authors.affiliation: Department of Biochemistry and Microbiology, Rutgers University, New Brunswick, New Jersey 08901, USA.
authors.forename: Maryam
authors.initials: M
authors.lastname: Honarbakhsh
authors.forename: Aramis A
authors.initials: AA
authors.lastname: Villafane
authors.forename: Ilona
authors.initials: I
authors.lastname: Ruhl
authors.forename: David
authors.initials: D
authors.lastname: Sannino
authors.forename: Elisabetta
authors.initials: E
authors.lastname: Bini
mesh_terms: D003001:Cloning, Molecular; D020869:Gene Expression Profiling; D017930:Genes, Reporter; D005966:Glucuronidase / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic; D055550:Protein Stability; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D048229:Sulfolobus solfataricus / Q000201:enzymology* / Q000235:genetics; D013696:Temperature
chemical_list: D011994:Recombinant Proteins; D005966:Glucuronidase