Nguyen V T, Morange M, Bensaude O
Département de Biologie Moléculaire, Institut Pasteur, Paris, France.
Anal Biochem. 1988 Jun;171(2):404-8. doi: 10.1016/0003-2697(88)90505-2.
The firefly enzyme luciferase catalyzes the luminescent reaction of luciferin with ATP and oxygen. The luciferase gene has recently been cloned and proposed as a reporter gene in procaryotic and eucaryotic cells. We present here a luciferase activity assay which relies on luminescence detection using a standard scintillation counter. This technique is simple, fast, inexpensive, and still very sensitive: as little as 0.02 pg (250,000 molecules) of enzyme is readily detected. The technique is optimized for the luciferase assay in mammalian cell lysates. Thus, the luciferase gene may become a very useful tool for gene regulation studies.
萤火虫酶荧光素酶催化荧光素与ATP和氧气的发光反应。荧光素酶基因最近已被克隆，并被提议作为原核细胞和真核细胞中的报告基因。我们在此介绍一种荧光素酶活性测定方法，该方法依赖于使用标准闪烁计数器进行发光检测。这项技术简单、快速、廉价，而且仍然非常灵敏：低至0.02 pg（250,000个分子）的酶很容易被检测到。该技术针对哺乳动物细胞裂解物中的荧光素酶测定进行了优化。因此，荧光素酶基因可能成为基因调控研究中非常有用的工具。
