Irla, Marta; Heggeset, Tonje M.; Naerdal, Ingemar; Paul, Lidia; Haugen, Tone; Le, Simone B.; Brautaset, Trygve; Wendisch, Volker F.: Genome-based genetic tool development for Bacillus methanolicus: theta- and rolling circle-replicating plasmids for inducible gene expression and application [...]. In: Frontiers in Microbiology. Jg.7. 2016
Inhalt
- Strains, Plasmids, and Primers
- Molecular Cloning
- Media and Cultivation Conditions
- β-Galactosidase (LacZ) Activity Assay
- Flow Cytometry
- Plasmid Stability
- Estimation of Copy Number by the Means of Droplet Digital PCR
- High Cell Density Fed-Batch Methanol Fermentation
- Measurement of Cadaverine and Amino Acids
- Detection of α-Amylase Activity
- Results
- Comparison of Different Replicons for Plasmid-Based Gene Expression in B. methanolicus MGA3
- Cadaverine Production from Methanol by Expression of a Heterologous Lysine Decarboxylase Gene from a Theta-Replicating Plasmid
- Plasmid Compatibility
- Construction of Mannitol Inducible Gene Expression System
- Xylose Inducible Gene Expression in B. methanolicus
- Introduction of Heterologous Starch Degradation Pathway in B. methanolicus MGA3 by Heterologous Overexpression of α-Amylase Gene from Streptomyces griseus IMRU3570
- Discussion
- Author Contributions
- Acknowledgments
- Supplementary Material
- References