Jungle Express is a versatile repressor system for tight transcriptional control

Ruegg, Thomas L.; Pereira, Jose H.; Chen, Joseph C.; DeGiovanni, Andy; Novichkov, Pavel; Mutalik, Vivek K.; Tomaleri, Giovani P.; Singer, Steven W.; Hillson, Nathan J.; Simmons, Blake A.; Adams, Paul D.; Thelen, Michael P.

doi:10.1038/s41467-018-05857-3

Title: Jungle Express is a versatile repressor system for tight transcriptional control

Journal Article · Thu Sep 06 00:00:00 EDT 2018 · Nature Communications

DOI:https://doi.org/10.1038/s41467-018-05857-3· OSTI ID:1467933

Ruegg, Thomas L.; Pereira, Jose H.; Chen, Joseph C.; DeGiovanni, Andy; Novichkov, Pavel; Mutalik, Vivek K.; Tomaleri, Giovani P.;

;

; Adams, Paul D.; Thelen, Michael P.

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.

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Research Organization:: Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Biological and Environmental Research (BER); USDOE Office of Science (SC), Basic Energy Sciences (BES); National Inst. of Health (NIH) (United States); Howard Hughes Medical Inst. (United States); Swiss National Science Foundation (SNSF)

Grant/Contract Number:: AC02-05CH11231; AC52-07NA27344; SC3GM096943

OSTI ID:: 1467933

Alternate ID(s):: OSTI ID: 1477413

Journal Information:: Nature Communications, Journal Name: Nature Communications Vol. 9 Journal Issue: 1; ISSN 2041-1723

Publisher:: Nature Publishing GroupCopyright Statement

Country of Publication:: United Kingdom

Language:: English

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Cited by: 26 works

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