PT  - JOURNAL ARTICLE
AU  - Huseyin Tas
AU  - Ángel Goñi-Moreno
AU  - Víctor de Lorenzo
TI  - A standardized broad host range inverter package for genetic circuitry design in Gram-negative bacteria
AID  - 10.1101/2020.07.14.202754
DP  - 2020 Jan 01
TA  - bioRxiv
PG  - 2020.07.14.202754
4099  - http://biorxiv.org/content/early/2020/07/15/2020.07.14.202754.short
4100  - http://biorxiv.org/content/early/2020/07/15/2020.07.14.202754.full
AB  - Genetically encoded logic gates, especially inverters—NOT gates—are the building blocks for designing circuits, engineering biosensors or decision-making devices in synthetic biology. However, the repertoire of inverters readily available for different species is rather limited. In this work, a large whole of NOT gates that was shown to function previously in a specific strain of Escherichia coli, was recreated as broad host range (BHR) collection of constructs assembled in low, medium and high copy number plasmid backbones of the SEVA (Standard European Vector Architecture) collection. The input/output function of each of the gates was characterized and parameterized in the environmental bacterium and metabolic engineering chassis Pseudomonas putida. Comparisons of the resulting fluorescence cytometry data with those published for the same gates in Escherichia coli provided useful hints on the portability of the corresponding gates. The hereby described BHR inverter package (20 different versions of 12 distinct gates) thus becomes a toolbox of choice for designing genetic circuitries in a variety of Gram-negative species other than E. coli.Competing Interest StatementThe authors have declared no competing interest.BHRbroad host rangeSDShine-Dalgarno sequenceRBSribosomal binding sequenceIPTGisopropyl thiogalactopyranosideCADcomputer-assisted designSEVAStandard European Vector Architecture