Abstract
The purpose of this study was to develop novel methods for attachment and cultivation of specifically positioned single yeast cells on a microelectrode surface with the application of a weak electrical potential. Saccharomyces cerevisiae diploid strains attached to an indium tin oxide/glass (ITO) electrode to which a negative potential between −0.2 and −0.4 V vs. Ag/AgCl was applied, while they did not adhere to a gallium-doped zinc oxide/glass electrode surface. The yeast cells attached to the negative potential-applied ITO electrodes showed normal cell proliferation. We found that the flocculin FLO10 gene-disrupted diploid BY4743 mutant strain (flo10Δ /flo10Δ) almost completely lost the ability to adhere to the negative potential-applied ITO electrode. Our results indicate that the mechanisms of diploid BY4743 S. cerevisiae adhesion involve interaction between the negative potential-applied ITO electrode and the Flo10 protein on the cell wall surface. A combination of micropatterning techniques of living single yeast cell on the ITO electrode and omics technologies holds potential of novel, highly parallelized, microchip-based single-cell analysis that will contribute to new screening concepts and applications.
Highlights
The first complete genome sequence of the yeast Saccharomyces cerevisiae was published (Goffeau et al 1996), enabling a wide variety of genetic engineering techniques and the strict functional analyses of proteins (Giaever et al 2002; Shibasaki, Maeda and Ueda 2009; Breker, Gymrek and Schuldiner 2013)
Saccharomyces cerevisiae diploid strains attached to an indium tin oxide/glass (ITO) electrode to which a negative potential between −0.2 and −0.4 V vs. Ag/AgCl was applied, while they did not adhere to a gallium-doped zinc oxide/glass electrode surface
Our results indicate that the mechanisms of diploid BY4743 S. cerevisiae adhesion involve interaction between the negative potential-applied ITO electrode and the Flo10 protein on the cell wall surface
Summary
The first complete genome sequence of the yeast Saccharomyces cerevisiae was published (Goffeau et al 1996), enabling a wide variety of genetic engineering techniques and the strict functional analyses of proteins (Giaever et al 2002; Shibasaki, Maeda and Ueda 2009; Breker, Gymrek and Schuldiner 2013). A heterozygous diploid mutant collection of approximately 6000 strains of S. cerevisiae, in each of which one copy of a single gene is deleted, is commercially available With this collection, it is possible to evaluate the role of each gene product in the response of cells to a drug (Lum et al 2004; Parsons et al 2004, 2006; Roberge 2008). Drug-induced haploinsufficiency profiling (HIP)/homozygous profiling (HOP) assay was one of the first assays to take advantage of parallelized growth strategy (Smith et al 2010). The HIP/HOP assay has been applied to identify the protein targets in numerous drugs and successfully employed in industry (Smith et al 2010; Giaever and Nislow 2014)
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