Szurmant Hendrik, Fukushima Tatsuya, Hoch James A
Department of Molecular and Structural Biochemistry, North Carolina State University, North Carolina, USA.
Methods Enzymol. 2007;422:396-417. doi: 10.1016/S0076-6879(06)22020-2.
The YycFG two-component system, highly conserved in the low G+C gram positives, is essential for cell viability in most organisms in which it has been studied. The system is organized within an operon that includes at least one but often three to four other genes. Products of two of these genes, yycH and yycI, have been shown to have a regulatory role on this two-component system. Immunofluorescent studies identified YycG kinase localization at the cell division sites consistent with its role in regulating cell divisional processes. The essential nature and operon organization of this system commanded special requirements in studying this system genetically. This chapter presents methods utilized in identifying the regulatory circuit that controls the activity of the YycG kinase in Bacillus subtilis. Most aspects of our approaches are applicable to other two-component systems in B. subtilis and the gram positives. Some are limited to essential systems, such as the YycFG system.
YycFG双组分系统在低G+C革兰氏阳性菌中高度保守，在大多数已研究的生物体中，该系统对细胞活力至关重要。该系统由一个操纵子组成，该操纵子至少包含一个，但通常包含三到四个其他基因。其中两个基因yycH和yycI的产物已被证明对这个双组分系统具有调节作用。免疫荧光研究确定YycG激酶定位于细胞分裂位点，这与其在调节细胞分裂过程中的作用一致。该系统的本质特性和操纵子组织对其进行遗传学研究提出了特殊要求。本章介绍了用于确定控制枯草芽孢杆菌中YycG激酶活性的调节回路的方法。我们方法的大多数方面适用于枯草芽孢杆菌和革兰氏阳性菌中的其他双组分系统。有些方法仅限于必需系统，例如YycFG系统。
