Journal
YEAST
Volume 34, Issue 8, Pages 343-355Publisher
WILEY
DOI: 10.1002/yea.3237
Keywords
Schizosaccharomyces; continuous culture; experimental evolution; chemostats; turbidostats
Categories
Funding
- NSF [1120425]
- NIH [P41 GM103533]
- ERC under the European Union's Seventh Framework Program (FP7)/ERC grant [310849]
- European Research Council (ERC) [310849] Funding Source: European Research Council (ERC)
Ask authors/readers for more resources
Maintenance of long-term cultures of yeast cells is central to a broad range of investigations, from metabolic studies to laboratory evolution assays. However, repeated dilutions of batch cultures lead to variations in medium composition, with implications for cell physiology. In Saccharomyces cerevisiae, powerful miniaturized chemostat setups, or ministat arrays, have been shown to allow for constant dilution of multiple independent cultures. Here we set out to adapt these arrays for continuous culture of a morphologically and physiologically distinct yeast, the fission yeast Schizosaccharomyces pombe, with the goal of maintaining constant population density over time. First, we demonstrated that the original ministats are incompatible with growing fission yeast for more than a few generations, prompting us to modify different aspects of the system design. Next, we identified critical parameters for sustaining unbiased vegetative growth in these conditions. This requires deletion of the gsf2 flocculin-encoding gene, along with addition of galactose to the medium and lowering of the culture temperature. Importantly, we improved the flexibility of the ministats by developing a piezo-pump module for the independent regulation of the dilution rate of each culture. This made it possible to easily grow strains that have different generation times in the same assay. Our system therefore allows for maintaining multiple fission yeast cultures in exponential growth, adapting the dilution of each culture over time to keep constant population density for hundreds of generations. These multiplex culture systems open the door to a new range of long-term experiments using thismodel organism. (C) 2017 TheAuthors. Yeast published by JohnWiley& Sons, Ltd.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available