ausblenden:
Schlagwörter:
Genetic Vectors/*genetics
Methylobacterium/*genetics/metabolism
*Operon
Recombinant Proteins/biosynthesis/genetics
*Transformation, Bacterial
Agrobacterium
Alphaproteobacteria
Caulobacter
Paracoccus
biobricks
operon assembly
promoter library
vector system
Zusammenfassung:
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.
