Characterization of a Fluorescent Protein Reporter System

Chemical and biological threats are ever present and attacks have occurred throughout the world in both war and peace-time. Multiple government agencies, academia, and private industry are developing detection capabilities to address such threats. The research presented in this paper supports development of a modular synthetic biologybased system that detects and reports the presence of a threat agent. Synthetic biology builds upon past research in genetic engineering and seeks to combine broad applications within biotechnology in novel ways. This basic research project will help to demonstrate a proof-of-concept design which will guide future studies on the development of a modular sensor platform. In this study, in vitro and in vivo techniques were used to evaluate a reporter system composed of the Tobacco Etch Virus (TEV) protease and an engineered fluorescent fusion protein for their combined effectiveness as a reporter system. The TEV protease demonstrated the ability to cleave the fusion protein construct to produce a measureable output signal. The coupling of this reporter system with a riboswitch recognition element showed that this system can be applied to detecting chemicals. Further studies to modify this recognition element suggest modularity for future biosensors to detect chemicals of military interest.

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