Abstract
Competitive growth between over 6,000 heterozygous yeast mutants in the presence of chromium together with microarray-based screens showed that proteasomal activity is crucial for cellular chromium resistance.
Highlights
The serious biological consequences of metal toxicity are well documented, but the key modes of action of most metals are unknown
Metal toxicity is widely studied in microorganisms, both as models to further our understanding of cellular metal toxicology, and because of the importance of metal toxicity in microbial biotechnologies [4,5,6,7]
Identification of heterozygotes with altered chromium resistances The experimental system involved co-culture of >6,000 heterozygous deletion strains in carbon-limited continuous culture
Summary
The serious biological consequences of metal toxicity are well documented, but the key modes of action of most metals are unknown. This reflects (in part) the more subtle phenotypes that the reduction in the copy number of a given gene (from two copies to one), as opposed to its complete removal, is expected to produce This subtlety means that small differences in growth rate of individual heterozygous mutants must be detected, and this is most achieved by competition experiments. Total genomic DNA extracted from the mixed competitions is subjected to PCR with the universal primers, yielding a pool of amplified tag sequences in which the abundance of each unique tag corresponds to the abundance of a strain in the culture [14,16] These abundances can be determined quantitatively by hybridization to oligonucleotide arrays, the data revealing the relative growth of each yeast mutant under the growth condition(s) of interest
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