Optical Sensing in Cell-Free Expression

Abstract

Light can be used as a control switch for gene expression with potential advantages, avoiding the defects induced by chemical substances. By transplanting components capable of emitting light at a specific wavelength from cells into a cell-free synthesis system, controlled gene expression can be achieved in vitro. Here, we describe an effective method to achieve optical sensing in cell-free protein synthesis (CFPS) based on Escherichia coli crude extract containing the two-component system (TCSs) YF1/FixJ, which was able to respond to blue light. Plasmids capable of interacting with the photosensitive components were constructed, and the fluorescent protein mCherry was used as a reporter. This protocol provides a detailed procedure guiding how to construct the blue-light sensing system in CFPS.

Keywords: Cell-free protein synthesis; Optical sensing; Two-component system.