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dc.contributor.authorEnglaender, Jacob A.
dc.contributor.authorJones, J. Andrew
dc.contributor.authorCress, Brady F.
dc.contributor.authorKuhlman, Thomas E.
dc.contributor.authorLinhardt, Robert J.
dc.contributor.authorKoffas, Mattheos A.G.
dc.identifier.citationEffect of Genomic Integration Location on Heterologous Protein Expression and Metabolic Engineering in E. coli. J. Englaender, A. J. Jones, B. Cress, T. Kuhlman, R. J. Linhardt, M.A.G. Koffas, ACS Synthetic Biology, 6, 710−720, 2017.
dc.descriptionACS Synthetic Biology, 6, 710−720
dc.descriptionNote : if this item contains full text it may be a preprint, author manuscript, or a Gold OA copy that permits redistribution with a license such as CC BY. The final version is available through the publisher’s platform.
dc.description.abstractChromosomal integration offers a selection-free alternative to DNA plasmids for expression of foreign proteins and metabolic pathways. Episomal plasmid DNA is convenient but has drawbacks including increased metabolic burden and the requirement for selection in the form of antibiotics. E. coli has long been used for the expression of foreign proteins and for the production of valuable metabolites by expression of complete metabolic pathways. The gene encoding the fluorescent reporter protein mCherry was integrated into four genomic loci on the E. coli chromosome to measure protein expression at each site. Expression levels ranged from 25% to 500% compared to the gene expressed on a high-copy plasmid. Modular expression of DNA is one of the most commonly used methods for optimizing metabolite production by metabolic engineering. By combining a recently developed method for integration of large synthetic DNA constructs into the genome, we were able to integrate two foreign pathways into the same four genomic loci. We have demonstrated that only one of the genomic loci resulted in the production of violacein, and that all four loci produced trans-cinnamic acid from the TAL pathway.
dc.description.sponsorshipNational Science Foundation
dc.relation.ispartofThe Linhardt Research Labs Online Collection
dc.relation.ispartofRensselaer Polytechnic Institute, Troy, NY
dc.relation.ispartofACS Synthetic Biology
dc.subjectChemistry and chemical biology
dc.subjectChemical and biological engineering
dc.subjectBiomedical engineering
dc.titleEffect of Genomic Integration Location on Heterologous Protein Expression and Metabolic Engineering in E. coli
dc.rights.holderIn Copyright : this Item is protected by copyright and/or related rights. You are free to use this Item in any way that is permitted by the copyright and related rights legislation that applies to your use. For other uses you need to obtain permission from the rights-holder(s).
dc.relation.departmentThe Linhardt Research Labs.
dc.relation.departmentThe Shirley Ann Jackson, Ph.D. Center for Biotechnology and Interdisciplinary Studies (CBIS)

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