An easily synthesized, photolyzable luciferase substrate for in vivo luciferase activity measurement.
An easily synthesized, photolyzable luciferase substrate for in vivo luciferase activity measurement.
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DOI：
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发表时间：
1993-11
期刊：
影响因子：
2.7
通讯作者：
Jiwei Yang;D. Thomason
中科院分区：
文献类型：
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作者：
Jiwei Yang;D. Thomason
许多报告基因测定需要通过固定或裂解来杀死细胞。对于活细胞中的测定，底物输送效率低下，并且不能以推注形式原位供应，这使得测定具有很大的可变性。我们报告了一种荧光素酯的简单合成方法，该酯既可光解又可被内源酯酶裂解，从而可以轻松监测活细胞中的荧光素酶活性。尽管光解底物可以以大剂量形式递送，但细胞中荧光素酯的快速平衡和内源酯酶的连续递送允许稳定、长期地测量荧光素酶活性。
Many reporter gene assays require killing the cell by fixation or lysis. For assays in living cells, the substrate delivery is inefficient and cannot be supplied in situ in a bolus, which makes assays highly variable. We report a simple synthesis of a luciferin ester that is both photolyzable and cleaved by endogenous esterases such that luciferase activity in living cells is easily monitored. Although the photolyzed substrate can be delivered in bolus, the rapid equilibration of the luciferin ester in the cell and the continuous delivery by the endogenous esterases allow stable, long-term measurements of luciferase activity.