Jungle Express is a versatile repressor system for tight transcriptional control

Thomas L. Ruegg, Jose H. Pereira, Joseph C. Chen, Andy DeGiovanni, Pavel Novichkov, Vivek K. Mutalik, Giovani P. Tomaleri, Steven W. Singer, Nathan J. Hillson, Blake A. Simmons, Paul D. Adams and Michael P. Thelen ()
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Thomas L. Ruegg: Joint BioEnergy Institute
Jose H. Pereira: Joint BioEnergy Institute
Joseph C. Chen: Joint BioEnergy Institute
Andy DeGiovanni: Joint BioEnergy Institute
Pavel Novichkov: Lawrence Berkeley National Laboratory
Vivek K. Mutalik: Lawrence Berkeley National Laboratory
Giovani P. Tomaleri: Joint BioEnergy Institute
Steven W. Singer: Joint BioEnergy Institute
Nathan J. Hillson: Joint BioEnergy Institute
Blake A. Simmons: Joint BioEnergy Institute
Paul D. Adams: Joint BioEnergy Institute
Michael P. Thelen: Joint BioEnergy Institute

Nature Communications, 2018, vol. 9, issue 1, 1-13

Abstract: Abstract Tightly regulated promoters are essential for numerous biological applications, where strong inducibility, portability, and scalability are desirable. Current systems are often incompatible with large-scale fermentations due to high inducer costs and strict media requirements. Here, we describe the bottom-up engineering of ‘Jungle Express’, an expression system that enables efficient gene regulation in diverse proteobacteria. This system is guided by EilR, a multidrug-binding repressor with high affinity to its optimized operator and cationic dyes that act as powerful inducers at negligible costs. In E. coli, the engineered promoters exhibit minimal basal transcription and are inducible over four orders of magnitude by 1 µM crystal violet, reaching expression levels exceeding those of the strongest current bacterial systems. Further, we provide molecular insights into specific interactions of EilR with its operator and with two inducers. The versatility of Jungle Express opens the way for tightly controlled and efficient gene expression that is not restricted to host organism, substrate, or scale.

Date: 2018
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DOI: 10.1038/s41467-018-05857-3

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