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Nat Biotechnol. 2014 Dec;32(12):1276-81. doi: 10.1038/nbt.3053. Epub 2014 Nov 17.

Tunable protein degradation in bacteria.

Author information

1
1] Howard Hughes Medical Institute, Boston University, Boston, Massachusetts, USA. [2] Center of Synthetic Biology, Boston University, Boston, Massachusetts, USA. [3] Department of Biomedical Engineering, Boston University, Boston, Massachusetts, USA.
2
1] Howard Hughes Medical Institute, Boston University, Boston, Massachusetts, USA. [2] Center of Synthetic Biology, Boston University, Boston, Massachusetts, USA. [3] Department of Biomedical Engineering, Boston University, Boston, Massachusetts, USA. [4] Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, Massachusetts, USA.

Abstract

Tunable control of protein degradation in bacteria would provide a powerful research tool. Here we use components of the Mesoplasma florum transfer-messenger RNA system to create a synthetic degradation system that provides both independent control of steady-state protein level and inducible degradation of targeted proteins in Escherichia coli. We demonstrate application of this system in synthetic circuit development and control of core bacterial processes and antibacterial targets, and we transfer the system to Lactococcus lactis to establish its broad functionality in bacteria. We create a 238-member library of tagged essential proteins in E. coli that can serve as both a research tool to study essential gene function and an applied system for antibiotic discovery. Our synthetic protein degradation system is modular, does not require disruption of host systems and can be transferred to diverse bacteria with minimal modification.

PMID:
25402616
PMCID:
PMC4262603
DOI:
10.1038/nbt.3053
[Indexed for MEDLINE]
Free PMC Article

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