Development of a thermostable β-glucuronidase-based reporter system for monitoring gene expression in hyperthermophiles
| Autor: | Aramis A. Villafane, Elisabetta Bini, David Sannino, Maryam Honarbakhsh, Ilona Ruhl |
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| Rok vydání: | 2012 |
| Předmět: |
ved/biology.organism_classification_rank.species
Bioengineering Applied Microbiology and Biotechnology Genes Reporter Gene expression Cloning Molecular Promoter Regions Genetic Gene Glucuronidase Reporter gene biology Protein Stability ved/biology Gene Expression Profiling Thermophile Sulfolobus solfataricus Temperature biology.organism_classification Molecular biology Recombinant Proteins Hyperthermophile Sulfolobus Biochemistry Biotechnology |
| Zdroj: | Biotechnology and Bioengineering. 109:1881-1886 |
| ISSN: | 0006-3592 |
| DOI: | 10.1002/bit.24432 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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