Development of a thermostable β‐glucuronidase‐based reporter system for monitoring gene expression in hyperthermophiles
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. Biotechnol. Bioeng. 2012; 109:1881–1886. © 2012 Wiley Periodicals, Inc.
Document Type: Research Article
Affiliations: Department of Biochemistry and Microbiology, Rutgers University, New Brunswick, New Jersey 08901; telephone: +1-732-932-9763 ext.122;, Fax: +1-732-932-8965
Publication date: 2012-07-01