Loading...
A set of synthetic versatile genetic control elements for the efficient expression of genes in Actinobacteria.
Horbal, Lilya Siegl, Theresa
Horbal, Lilya
Siegl, Theresa
Citations
Altmetric:
Advisors
Editors
Other Contributors
Issue Date
2018-01-11
Submitted date
Files
Loading...
Open Access publication
Adobe PDF, 1.72 MB
Other Titles
Abstract
The design and engineering of secondary metabolite gene clusters that are characterized by complicated genetic organization, require the development of collections of well-characterized genetic control elements that can be reused reliably. Although a few intrinsic terminators and RBSs are used routinely, their translation and termination efficiencies have not been systematically studied in Actinobacteria. Here, we analyzed the influence of the regions surrounding RBSs on gene expression in these bacteria. We demonstrated that inappropriate RBSs can reduce the expression efficiency of a gene to zero. We developed a genetic device - an in vivo RBS-selector - that allows selection of an optimal RBS for any gene of interest, enabling rational control of the protein expression level. In addition, a genetic tool that provides the opportunity for measurement of termination efficiency was developed. Using this tool, we found strong terminators that lead to a 17-100-fold reduction in downstream expression and are characterized by sufficient sequence diversity to reduce homologous recombination when used with other elements. For the first time, a C-terminal degradation tag was employed for the control of protein stability in Streptomyces. Finally, we describe a collection of regulatory elements that can be used to control metabolic pathways in Actinobacteria.
Citation
A set of synthetic versatile genetic control elements for the efficient expression of genes in Actinobacteria. 2018, 8 (1):491 Sci Rep
Publisher
Journal
PubMed ID
PubMed Central ID
Additional Links
Embedded video
Type
Article
Language
en
Description
Series/Report no.
ISSN
2045-2322