A set of synthetic versatile genetic control elements for the efficient expression of genes in Actinobacteria.
Cast your vote
You can rate an item by clicking the amount of stars they wish to award to this item.
When enough users have cast their vote on this item, the average rating will also be shown.
Your vote was cast
Thank you for your feedback
Thank you for your feedback
MetadataShow full item record
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.
CitationA set of synthetic versatile genetic control elements for the efficient expression of genes in Actinobacteria. 2018, 8 (1):491 Sci Rep
AffiliationHIPS, Helmholtz-Institut für pharmazeutische Forschung Saarland, Universitätscampus 8.1, 66123 Saarbrücken, Germany.
The following license files are associated with this item:
Except where otherwise noted, this item's license is described as http://creativecommons.org/licenses/by-nc-sa/4.0/
- Translation efficiency of heterologous proteins is significantly affected by the genetic context of RBS sequences in engineered cyanobacterium Synechocystis sp. PCC 6803.
- Authors: Thiel K, Mulaku E, Dandapani H, Nagy C, Aro EM, Kallio P
- Issue date: 2018 Mar 2
- Measurement and modeling of intrinsic transcription terminators.
- Authors: Cambray G, Guimaraes JC, Mutalik VK, Lam C, Mai QA, Thimmaiah T, Carothers JM, Arkin AP, Endy D
- Issue date: 2013 May
- Influence of transcriptional and translational control sequences on the expression of foreign genes in Caulobacter crescentus.
- Authors: Yap WH, Thanabalu T, Porter AG
- Issue date: 1994 May
- Functional genetic elements for controlling gene expression in <i>Cupriavidus necator</i> H16.
- Authors: Alagesan S, Hanko EKR, Malys N, Ehsaan M, Winzer K, Minton NP
- Issue date: 2018 Jul 20
- Comparative analysis of RNA regulatory elements of amino acid metabolism genes in Actinobacteria.
- Authors: Seliverstov AV, Putzer H, Gelfand MS, Lyubetsky VA
- Issue date: 2005 Oct 3