用于高通量功能筛选应用的海肾荧光素酶变体。
Gaussia luciferase variant for high-throughput functional screening applications.
机构信息
Molecular Neurogenetics Unit, Department of Neurology, Harvard Medical School, Boston Massachusetts 02115, USA.
出版信息
Anal Chem. 2009 Aug 15;81(16):7102-6. doi: 10.1021/ac901234r.
Abstract
Gaussia luciferase (Gluc) is a sensitive reporter for studying different biological processes such as gene expression, promoter activity, protein-protein interactions, signal transduction, as well as tumor cell growth and response to therapy. Since Gluc is naturally secreted, the kinetics of these processes can be monitored in real-time by measuring an aliquot of conditioned medium in culture or a few microliters of blood in vivo at different time points. Gluc catalyzes light emission with a short half-life which is unfavorable for certain applications. We isolated a Gluc mutant that catalyzes enhanced light stability in the presence of a detergent, in combination with high sensitivity, making it an attractive luciferase for high-throughput functional screening applications.
摘要
海肾荧光素酶（Gluc）是一种灵敏的报告基因，可用于研究多种生物学过程，如基因表达、启动子活性、蛋白质-蛋白质相互作用、信号转导以及肿瘤细胞的生长和对治疗的反应等。由于 Gluc 是天然分泌的，因此可以通过在不同时间点测量培养物中的条件培养基的等分试样或体内的几微升血液，实时监测这些过程的动力学。Gluc 催化的发光具有半衰期短的缺点，不利于某些应用。我们分离出一种 Gluc 突变体，该突变体在去污剂存在下催化增强的光稳定性，同时保持高灵敏度，使其成为高通量功能筛选应用的有吸引力的荧光素酶。
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