Robertson J Brian, Johnson Carl Hirschie
Department of Biological Sciences, Vanderbilt University, Nashville, TN, USA.
Methods Mol Biol. 2011;734:63-79. doi: 10.1007/978-1-61779-086-7_4.
This chapter describes a method for generating yeast respiratory oscillations in continuous culture and monitoring rhythmic promoter activity of the culture by automated real-time recording of luminescence. These techniques chiefly require the use of a strain of Saccharomyces cerevisiae that has been genetically modified to express firefly luciferase under the control of a promoter of interest and a continuous culture bioreactor that incorporates a photomultiplier apparatus for detecting light emission. Additionally, this chapter describes a method for observing rhythmic (cell cycle-related) promoter activity in small batch cultures of yeast through luminescence monitoring.
本章介绍了一种在连续培养中产生酵母呼吸振荡并通过自动实时记录发光来监测培养物节律性启动子活性的方法。这些技术主要需要使用经过基因改造的酿酒酵母菌株，使其在感兴趣的启动子控制下表达萤火虫荧光素酶，以及一个连续培养生物反应器，该反应器配备有用于检测光发射的光电倍增装置。此外，本章还描述了一种通过发光监测在酵母小批量培养中观察节律性（与细胞周期相关）启动子活性的方法。
