1887

Abstract

Synthetic genes encoding functional luciferases of the click beetle (CB) have been expressed in the human fungal pathogen Both green- and red-emitting CB luciferases (CaCBGluc and CaCBRluc) were produced with high efficiency in transformants under transcriptional control of the growth-dependent promoter, as well as by the and promoters, which are upregulated during hyphal morphogenesis, as well as by theand promoters, which are downregulated. For all hyphally regulated genes, relative bioluminescence values derived from promoter fusions approximated relative transcript levels of native genes, although downregulation of promoter activity required correction for the stability of CB luciferases (approximate half-lives 30 min for CaCBRluc and 80 min for CaCBGluc, as determined by immunoblotting). Importantly, the activity of both luciferases could be separately monitored in a single strain, in intact cells, in lysed cells or in cell extracts using luciferin as single substrate and inhibition of hypha formation by farnesol could be easily detected by the fusion. The results suggest that CB luciferases are convenient tools to measure gene expression in and may facilitate screenings for antifungal compounds.

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2016-08-01
2022-01-29
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