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Development of a thermostable β‐glucuronidase‐based reporter system for monitoring gene expression in hyperthermophiles
Author(s) -
Honarbakhsh Maryam,
Villafane Aramis A.,
Ruhl Ilona,
Sannino David,
Bini Elisabetta
Publication year - 2012
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.24432
Subject(s) - hyperthermophile , sulfolobus , sulfolobus solfataricus , thermophile , reporter gene , glucuronidase , gene , biology , gene expression , biochemistry , enzyme , archaea
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 solfataricu s. 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.