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Bicistronic Design-Based Continuous and High-Level Membrane Protein Production in Escherichia coil
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Number of Authors: 82019 (English)In: ACS Synthetic Biology, E-ISSN 2161-5063, Vol. 8, no 7, p. 1685-1690Article in journal (Refereed) Published
Abstract [en]

Escherichia coli has been widely used as a platform microorganism for both membrane protein production and cell factory engineering. The current methods to produce membrane proteins in this organism require the induction of target gene expression and often result in unstable, low yields. Here, we present a method combining a constitutive promoter with a library of bicistronic design (BCD) elements, which enables inducer-free, tuned translation initiation for optimal protein production. Our system mediates stable, constitutive production of bacterial membrane proteins at yields that outperform those obtained with E. coli Lemo21(DE3), the current gold standard for bacterial membrane protein production. We envisage that the continuous, fine-tunable, and high-level production of membrane proteins by our method will greatly facilitate their study and their utilization in engineering cell factories.

Place, publisher, year, edition, pages
2019. Vol. 8, no 7, p. 1685-1690
Keywords [en]
protein production, membrane proteins, translational coupling, bicistronic design, Escherichia coli
National Category
Biological Sciences
Identifiers
URN: urn:nbn:se:su:diva-171792DOI: 10.1021/acssynbio.9b00101ISI: 000476957300024PubMedID: 31264406OAI: oai:DiVA.org:su-171792DiVA, id: diva2:1346154
Available from: 2019-08-27 Created: 2019-08-27 Last updated: 2019-08-27Bibliographically approved

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Finger-Bou, Maxde Gier, Jan-Willem
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