A set of versatile brick vectors and promoters for the assembly, expression, 
and integration of synthetic operons in Methylobacterium extorquens AM1 and 
other alphaproteobacteria

Schada von Borzyskowski, L.; Remus-Emsermann, M.; Weishaupt, R.; Vorholt, J. A.; Erb, Tobias J.

doi:10.1021/sb500221v

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A set of versatile brick vectors and promoters for the assembly, expression, and integration of synthetic operons in Methylobacterium extorquens AM1 and other alphaproteobacteria

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https://www.ncbi.nlm.nih.gov/pubmed/25105793
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Citation

Schada von Borzyskowski, L., Remus-Emsermann, M., Weishaupt, R., Vorholt, J. A., & Erb, T. J. (2014). A set of versatile brick vectors and promoters for the assembly, expression, and integration of synthetic operons in Methylobacterium extorquens AM1 and other alphaproteobacteria. ACS Synth Biol, 4(4), 430-43. doi:10.1021/sb500221v.

Cite as: https://hdl.handle.net/21.11116/0000-000A-CAFB-8

Abstract

The discipline of synthetic biology requires standardized tools and genetic elements to construct novel functionalities in microorganisms; yet, many model systems still lack such tools. Here, we describe a novel set of vectors that allows the convenient construction of synthetic operons in Methylobacterium extorquens AM1, an important alphaproteobacterial model organism for methylotrophy and a promising platform organism for methanol-based biotechnology. In addition, we provide a set of constitutive alphaproteobacterial promoters of different strengths that were characterized in detail by two approaches: on the single-cell scale and on the cell population level. Finally, we describe a straightforward strategy to deliver synthetic constructs to the genome of M. extorquens AM1 and other Alphaproteobacteria. This study defines a new standard to systematically characterize genetic parts for their use in M. extorquens AM1 by using single-cell fluorescence microscopy and opens the toolbox for synthetic biological applications in M. extorquens AM1 and other alphaproteobacterial model systems.