Department of Biochemistry, University of Bayreuth, Bayreuth, Germany.
Department of Anatomy, University of Helsinki, Helsinki, Finland.
Methods Mol Biol. 2024;2760:463-477. doi: 10.1007/978-1-0716-3658-9_26.
By applying sensory photoreceptors, optogenetics realizes the light-dependent control of cellular events and state. Given reversibility, noninvasiveness, and exquisite spatiotemporal precision, optogenetic approaches enable innovative use cases in cell biology, synthetic biology, and biotechnology. In this chapter, we detail the implementation of the pREDusk, pREDawn, pCrepusculo, and pAurora optogenetic circuits for controlling bacterial gene expression by red and blue light, respectively. The protocols provided here guide the practical use and multiplexing of these circuits, thereby enabling graded protein production in bacteria at analytical and semi-preparative scales.
通过应用感觉光感受器,光遗传学实现了对细胞事件和状态的光依赖性控制。鉴于其可逆性、非侵入性和精细的时空精度,光遗传学方法在细胞生物学、合成生物学和生物技术中实现了创新的应用案例。在本章中,我们详细介绍了 pREDusk、pREDawn、pCrepusculo 和 pAurora 光遗传学回路的实施情况,这些回路分别通过红光和蓝光控制细菌基因表达。这里提供的方案指导了这些回路的实际使用和复用,从而能够在分析和半制备规模上对细菌进行分级蛋白质生产。