Scully Audra L, Zelhof Andrew C, Kay Steve A
Department of Cell Biology, The Scripps Research Institute, La Jolla, California 92037, USA.
Mol Cell Neurosci. 2002 Apr;19(4):501-14. doi: 10.1006/mcne.2001.1091.
Elucidating the mechanisms that link the circadian pacemaker to the timing of behaviors it controls is one of the greatest challenges in circadian biology. We report the generation of a P element, Pluc+, containing a novel reporter fusion. Our fusion reporter capitalizes on the use of luciferase bioluminescence to easily analyze temporal expression as well as the strength of myc epitopes and GFP to identify spatial expression. Using Pluc+ we have identified and characterized a novel C2H2 zinc-finger gene, regular (rgr), that cycles circadianly in phase with period (per) gene expression, but shifts to light-dark regulation in Clk(Jrk) mutant flies. By following myc expression of the Pluc+ reporter, we demonstrate that Rgr is expressed in a discrete number of neurons in the brain which overlap with axons expressing pigment-dispersing factor.
阐明将昼夜节律起搏器与其所控制行为的时间联系起来的机制，是昼夜节律生物学面临的最大挑战之一。我们报告了一个P因子Pluc +的产生，它包含一个新型报告基因融合体。我们的融合报告基因利用荧光素酶生物发光来轻松分析时间表达，以及利用myc表位和绿色荧光蛋白的强度来确定空间表达。使用Pluc +，我们鉴定并表征了一个新型C2H2锌指基因，即规律基因（rgr），它与周期基因（per）的表达同步进行昼夜节律循环，但在Clk（Jrk）突变果蝇中转变为光暗调节。通过追踪Pluc +报告基因的myc表达，我们证明Rgr在大脑中离散数量的神经元中表达，这些神经元与表达色素分散因子的轴突重叠。
