Department of Biochemistry and Microbiology, Rutgers University, New Brunswick, New Jersey 08901, USA.
Biotechnol Bioeng. 2012 Jul;109(7):1881-6. doi: 10.1002/bit.24432. Epub 2012 Jan 11.
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.
嗜中温葡糖醛酸酶是在植物中最广泛应用的基因表达报告酶，但由于其热稳定性不足，不适合在（超）嗜热菌中作为报告酶。在这里，我们介绍了源自嗜热硫磺酸杆菌的天然 66.8 kDa 耐热β-葡糖苷酸酶。该酶在很宽的温度范围内具有活性，非常适合但不仅限于对嗜热菌进行体内遗传研究。作为概念验证，我们通过监测与β-葡糖苷酸酶编码基因 gusB 和铜响应启动子融合创建的启动子融合，证明了其在 Sulfolobus 中作为基因表达报告酶的用途。
