Nakajima Yoshihiro, Kimura Takuma, Sugata Kazunori, Enomoto Toshiteru, Asakawa Atsushi, Kubota Hidehiro, Ikeda Masaaki, Ohmiya Yoshihiro
National Institute of Advanced Industrial Science and Technology, Osaka, Japan.
Biotechniques. 2005 Jun;38(6):891-4. doi: 10.2144/05386ST03.
Reporter assays that use luciferase are widely employed for monitoring cellular events associated with gene expression. In general, firefly luciferase and Renilla luciferase are used for monitoring single gene expression. However, the expression of more than one gene cannot be monitored simultaneously by this system because one of the two reporting luciferases must be used as an internal control. We have developed a novel reporter assay system in which three luciferases that emit green, orange, and red light with a single substrate are used as reporter genes. The activities of the luciferases can be measured simultaneously and quantitatively with optical filters. This system enables us to simply and rapidly monitor multiple gene expressions in a one-step reaction.
使用荧光素酶的报告基因检测法被广泛用于监测与基因表达相关的细胞事件。一般来说，萤火虫荧光素酶和海肾荧光素酶用于监测单个基因的表达。然而，该系统无法同时监测多个基因的表达，因为两种报告荧光素酶之一必须用作内对照。我们开发了一种新型报告基因检测系统，其中三种能利用单一底物发出绿色、橙色和红色光的荧光素酶被用作报告基因。通过光学滤光片可以同时且定量地测量荧光素酶的活性。该系统使我们能够在一步反应中简单快速地监测多个基因的表达。
