RT Journal Article
SR Electronic
T1 Parameterization of regulatory nodes for engineering broad host range heterologous gene expression
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2021.05.27.445938
DO 10.1101/2021.05.27.445938
A1 Pablo I. Nikel
A1 Ilaria Benedetti
A1 Victor de Lorenzo
A1 Belen Calles
YR 2021
UL http://biorxiv.org/content/early/2021/05/27/2021.05.27.445938.abstract
AB By building on the SEVA (Standard European Vector Architecture) format we have refactored a number of regulatory nodes recruited from both Gram-negative and Gram-positive bacteria for rigorously comparing and parameterizing five expression devices that respond to diverse and unrelated chemical inducers, i.e. LacIq-Ptrc, XylS-Pm, AlkS-PalkB, CprK-PDB3 and ChnR-PchnB. These were assembled as cargoes following the SEVA standard within exactly the same vector backbone and bearing the different functional segments arrayed in an invariable DNA scaffold. Their performance in an Escherichia coli strain of reference was then analyzed through the readout a fluorescence reporter gene that contained strictly identical translation signal elements in all cases and in the same DNA context. This study allowed us to describe and compare the cognate expression systems with unprecedented quantitative detail. The systems under scrutiny diverged considerably in their capacity, expression noise, inducibility and OFF/ON ratios. These features, along with the absence of physiological effects caused by the inducers and the lack of cross-regulation offer a panoply of choices to potential users and help interoperability of the specific constructs.Competing Interest StatementThe authors have declared no competing interest.