Optimized Origin for Improved Plasmid Maintenance Across Yeast Species Including P. pastoris
Principal Investigator: Maitreya Dunham
Therefore, there is a large unmet need in both research and industrial settings to generate plasmid backbones that are able to efficiently replicate and shuttle between yeast species. Additionally, there are some yeast species in which commercially available ARS sequences are unavailable. Due to this technical limitation, expression cassettes must be integrated, rendering large screens difficult or impossible in some cases. For example, the commercially relevant yeast strain Pichia pastoris, which is used in the biotech industry as a protein expression system due to its desirable properties, currently lacks a robust plasmid expression system.
Drs. Liachko and Dunham from the UW Department of Genome Sciences identified a DNA sequence that functions as an ARS across different yeast species. This sequence was further optimized to improve its function across multiple yeast species. Furthermore, the synthetic DNA sequence conferred stable plasmid replication function in all budding yeast species tested. Additionally, the investigators have identified and optimized ARSs that lead to efficient and stable plasmid propagation in P. pastoris which would allow for more efficient testing of protein constructs.
• Synthetic sequences function in at least 10 budding yeast species with diverse requirements.
• Robust plasmid expression stability in P. pastoris .
• Optimized sequences lead to more robust expression as compared to wild-type sequences.
For more info, contact: Andrea Valladao
- Liachko I, Dunham MJ (March 2014), An autonomously replicating sequence for use in a wide range of budding yeasts., FEMS Yeast Res.
- Liachko I, Youngblood RA, Tsui K, Bubb KL, Queitsch C, Raghuraman MK, Nislow C, Brewer BJ, Dunham MJ (3/6/2014), GC-rich DNA elements enable replication origin activity in the methylotrophic yeast Pichia pastoris., PLoS Genet.
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