Iijima-Ando Kanae, Yin Jerry C P
Cold Spring Harbor Laboratory, Cold Spring Harbor, New York 11724, USA.
Methods Enzymol. 2005;393:302-15. doi: 10.1016/S0076-6879(05)93013-9.
The cAMP response Element (CRE)-binding protein (CREB) is involved in many adaptive behaviors, including circadian rhythms. In order to assess CREB activity in vivo, we made transgenic flies carrying a CRE-luciferase reporter and showed that this reporter is CRE and dCREB2 responsive. dCREB2 is the Drosophila homolog of mammalian CREB?CREM. The transgenic luciferase activity cycles with a 24-h periodicity, suggesting that dCREB2 and period are somehow linked. The CRE-luciferase reporter is a useful monitor of circadian activity, and mutations can be found that affect its periodicity, baseline activity, or amplitude. Analysis of such mutations should reveal information about how particular genes affect the molecular machinery of circadian cycling and how different genes affect the activity of dCREB2.
环磷酸腺苷反应元件(CRE)结合蛋白(CREB)参与包括昼夜节律在内的多种适应性行为。为了评估体内的CREB活性,我们构建了携带CRE - 荧光素酶报告基因的转基因果蝇,并表明该报告基因对CRE和dCREB2有反应。dCREB2是哺乳动物CREB - CREM的果蝇同源物。转基因荧光素酶活性以24小时周期循环,表明dCREB2和周期基因在某种程度上存在联系。CRE - 荧光素酶报告基因是昼夜活动的有用监测指标,并且可以发现影响其周期性、基础活性或振幅的突变。对这些突变的分析应能揭示特定基因如何影响昼夜节律循环的分子机制以及不同基因如何影响dCREB2的活性。