Design and Control of Extrachromosomal Elements in Methylorubrum extorquens AM1
| Autor: | Florian Petit, Anke Becker, Martina Carrillo, Amelie Dransfeld, Marcel Wagner, Tobias J. Erb |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
DNA Replication
0106 biological sciences Inducible promoters Letter Extrachromosomal Inheritance Biomedical Engineering Computational biology Biology 01 natural sciences Biochemistry Genetics and Molecular Biology (miscellaneous) inducible promoters 03 medical and health sciences Synthetic biology Bacterial Proteins Methylobacterium extorquens 010608 biotechnology Extrachromosomal DNA Escherichia coli Promoter Regions Genetic Organism Alphaproteobacteria 030304 developmental biology 0303 health sciences DNA Helicases Methylorubrum extorquens Gene Expression Regulation Bacterial General Medicine repABC biology.organism_classification synthetic chromosomes Metabolic Engineering Trans-Activators Replicon Synthetic Biology Genome Bacterial Biotechnology Plasmids |
| Zdroj: | ACS Synthetic Biology |
| ISSN: | 2161-5063 |
| DOI: | 10.1021/acssynbio.9b00220 |
| Popis: | Genetic tools are a prerequisite to engineer cellular factories for synthetic biology and biotechnology. Methylorubrum extorquens AM1 is an important platform organism of a future C1-bioeconomy. However, its application is currently limited by the availability of genetic tools. Here we systematically tested repABC regions to maintain extrachromosomal DNA in M. extorquens. We used three elements to construct mini-chromosomes that are stably inherited at single copy number and can be shuttled between Escherichia coli and M. extorquens. These mini-chromosomes are compatible among each other and with high-copy number plasmids of M. extorquens. We also developed a set of inducible promoters of wide expression range, reaching levels exceeding those currently available, notably the PmxaF-promoter. In summary, we provide a set of tools to control the dynamic expression and copy number of genetic elements in M. extorquens, which opens new ways to unleash the metabolic and biotechnological potential of this organism for future applications. |
| Databáze: | OpenAIRE |
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