2.50
Hdl Handle:
http://hdl.handle.net/10033/621311
Title:
A set of synthetic versatile genetic control elements for the efficient expression of genes in Actinobacteria.
Authors:
Horbal, Lilya; Siegl, Theresa; Luzhetskyy, Andriy ( 0000-0001-6497-0047 )
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.
Affiliation:
HIPS, Helmholtz-Institut für pharmazeutische Forschung Saarland, Universitätscampus 8.1, 66123 Saarbrücken, Germany.
Citation:
A set of synthetic versatile genetic control elements for the efficient expression of genes in Actinobacteria. 2018, 8 (1):491 Sci Rep
Journal:
Scientific reports
Issue Date:
11-Jan-2018
URI:
http://hdl.handle.net/10033/621311
DOI:
10.1038/s41598-017-18846-1
PubMed ID:
29323285
Type:
Article
Language:
en
ISSN:
2045-2322
Appears in Collections:
publications of the research group metabolic engineering of Actinomycetes ([HIPS]AMEG)

Full metadata record

DC FieldValue Language
dc.contributor.authorHorbal, Lilyaen
dc.contributor.authorSiegl, Theresaen
dc.contributor.authorLuzhetskyy, Andriyen
dc.date.accessioned2018-03-07T15:01:40Z-
dc.date.available2018-03-07T15:01:40Z-
dc.date.issued2018-01-11-
dc.identifier.citationA set of synthetic versatile genetic control elements for the efficient expression of genes in Actinobacteria. 2018, 8 (1):491 Sci Repen
dc.identifier.issn2045-2322-
dc.identifier.pmid29323285-
dc.identifier.doi10.1038/s41598-017-18846-1-
dc.identifier.urihttp://hdl.handle.net/10033/621311-
dc.description.abstractThe 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.en
dc.language.isoenen
dc.rights.urihttp://creativecommons.org/licenses/by-nc-sa/4.0/*
dc.titleA set of synthetic versatile genetic control elements for the efficient expression of genes in Actinobacteria.en
dc.typeArticleen
dc.contributor.departmentHIPS, Helmholtz-Institut für pharmazeutische Forschung Saarland, Universitätscampus 8.1, 66123 Saarbrücken, Germany.en
dc.identifier.journalScientific reportsen

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