TY - JOUR
T1 - Construction of a genetic multiplexer to toggle between chemosensory pathways in Escherichia coli
AU - Moon, Tae Seok
AU - Clarke, Elizabeth J.
AU - Groban, Eli S.
AU - Tamsir, Alvin
AU - Clark, Ryan M.
AU - Eames, Matthew
AU - Kortemme, Tanja
AU - Voigt, Christopher A.
N1 - Funding Information:
We thank Ala Trusina and Patrick Visperas for the initial work as part of the 2006 University of California, San Francisco iGEM team. We thank Adam Arkin (University of California, Berkeley), John Parkinson (The University of Utah), and Mark Goulian (University of Pennsylvania) for the materials and help with assays. DNA 2.0 graciously provided synthesis for the University of California, San Francisco iGEM project. C.A.V. was supported by the Pew and Packard Foundations, Office of Naval Research, National Institutes of Health ( EY016546 and AI067699 ), National Science Foundation (NSF) ( BES-0547637 ), and a Sandler Family Opportunity Award. C.A.V. and T.K. are part of the NSF Synthetic Biology Engineering Research Center. T.K. was supported by the Sloan Foundation, an NSF CAREER award, and a Sandler Family Award.
PY - 2011/2/18
Y1 - 2011/2/18
N2 - Many applications require cells to switch between discrete phenotypic states. Here, we harness the FimBE inversion switch to flip a promoter, allowing expression to be toggled between two genes oriented in opposite directions. The response characteristics of the switch are characterized using two-color cytometry. This switch is used to toggle between orthogonal chemosensory pathways by controlling the expression of CheW and CheW*, which interact with the Tar (aspartate) and Tsr* (serine) chemoreceptors, respectively. CheW* and Tsr* each contain a mutation at their protein-protein interface such that they interact with each other. The complete genetic program containing an arabinose-inducible FimE controlling CheW/CheW* (and constitutively expressed tar/tsr*) is transformed into an Escherichia coli strain lacking all native chemoreceptors. This program enables bacteria to swim toward serine or aspartate in the absence or in the presence of arabinose, respectively. Thus, the program functions as a multiplexer with arabinose as the selector. This demonstrates the ability of synthetic genetic circuits to connect to a natural signaling network to switch between phenotypes.
AB - Many applications require cells to switch between discrete phenotypic states. Here, we harness the FimBE inversion switch to flip a promoter, allowing expression to be toggled between two genes oriented in opposite directions. The response characteristics of the switch are characterized using two-color cytometry. This switch is used to toggle between orthogonal chemosensory pathways by controlling the expression of CheW and CheW*, which interact with the Tar (aspartate) and Tsr* (serine) chemoreceptors, respectively. CheW* and Tsr* each contain a mutation at their protein-protein interface such that they interact with each other. The complete genetic program containing an arabinose-inducible FimE controlling CheW/CheW* (and constitutively expressed tar/tsr*) is transformed into an Escherichia coli strain lacking all native chemoreceptors. This program enables bacteria to swim toward serine or aspartate in the absence or in the presence of arabinose, respectively. Thus, the program functions as a multiplexer with arabinose as the selector. This demonstrates the ability of synthetic genetic circuits to connect to a natural signaling network to switch between phenotypes.
KW - GFP
KW - NSF
KW - National Science Foundation
KW - RBS
KW - RFP
KW - green fluorescent protein
KW - red fluorescent protein
KW - ribosome binding site
UR - http://www.scopus.com/inward/record.url?scp=79251595456&partnerID=8YFLogxK
U2 - 10.1016/j.jmb.2010.12.019
DO - 10.1016/j.jmb.2010.12.019
M3 - Article
C2 - 21185306
AN - SCOPUS:79251595456
SN - 0022-2836
VL - 406
SP - 215
EP - 227
JO - Journal of Molecular Biology
JF - Journal of Molecular Biology
IS - 2
ER -