Genetic Transformation and Tool Development
The ability to genetically modify microorganisms is foundational to performing rational strain engineering, but most organisms are not transformable when first isolated. We are able to develop initial transformation methodologies and protocols for new organisms or improve upon existing ones. We can further develop high throughput tools for more rapid strain modification in these new organisms, including phage integrase-based genome integration tools for rapid screening of gene expression libraries and CRISPR-based genome editing tools for rapid, targeted gene deletion or gene regulation. Transposon mutant and overexpression libraries can also be created to allow high throughput phenotype screening or selections. Promoter libraries can be created to enable rational selection of gene expression levels for heterologous expression. The ABF has deep expertise in non-model organisms, including work with extremophilic organisms that thrive at elevated temperature and extreme pH, including both aerobes and strict anaerobes. We are able to improve transformation efficiency and developed genetic tools for diverse organisms to accelerate rational strain engineering. Development of new microbial platforms or further improving genetic tools in existing platform organisms will provide unique new chassis organisms for advanced bioprocessing to enable a bioeconomy.
References and Additional Information:
Elmore J, Furches A, Wolff G, Gorday K, and Guss AM. Development of a high efficiency integration system and promoter library for rapid modification of Pseudomonas putida KT2440. Metabolic Engineering Communications 5 (2017) 1–8. DOI: j.meteno.2017.04.001
Guss AM, Olson DG, Caiazza NC, Lynd LR. (2012) Dcm methylation is detrimental to plasmid transformation in Clostridium thermocellum. Biotechnol Biofuels 5(1):30.
Yaegashi, J., et al. (2017) Rhodosporidium toruloides: a new platform organism for conversion of lignocellulose into terpene biofuels and bioproducts. Biotechnol Biofuels 10: p. 241
Oak Ridge National Laboratory
Los Alamos National Laboratory
Sandia National Laboratories