Abstract
Every year, Enterotoxigenic Escherichia coli (ETEC), the most common form of traveler’s diarrhea, affects thousands of military personnel deployed overseas. The goal of this research was to engineer non-pathogenic E. coli to sense ETEC, respond to its presence, and package the non-pathogenic E. coli in a cellulose matrix to enable environmental detection of ETEC. Two plasmids were created: ‘sense-respond’; and ‘packaging’. The sense-respond plasmid detected autoinducer 2 (AI-2), a quorum sensing molecule created by most ETEC strains, by expressing LsrR which switches on the Lsr promoter. Activation of the Lsr promoter expresses superfolder green fluorescent protein (sfGFP), indicating the presence of ETEC. The packaging plasmid expresses a fusion protein consisting of curli fibers and cellulose binding domains. These modified surface proteins permit the bacteria to bind to cellulose, encapsulating the sense-response module. This genetically engineered machine could be deployed in both the internal and external environment to detect ETEC.
Footnotes
1) Office of Secretary of Defense, ARAP Program: Synthetic Biology for Military Environments (A. Bete, J. Carter, C. Davis, J. Dong, M. Herrmann, H. Jesse, D. McDonald, P. Menart, A. Poole, A. Smith, D. Carter, M. Carter, Dr. M. Goodson, Dr. C. Hung, Dr. N. Kelley-Loughnane, R. Krabacher, Dr. V. Varaljay, Dr. M. Carter, Dr. C. Metosh-Dickey, and Dr. C. O’Malley)
a) The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Competing Interests: The authors have declared that no competing interests exist.
Ethics Statement: N/A
Data Availability: Yes – all data are fully available without restriction. All relevant data is within the paper and all other data can be found at http://2017.igem.org/Team:US_A…