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Luciferase NanoLuc as a reporter for gene expression and protein levels in Saccharomyces cerevisiae.

ABSTRACT: Reporter proteins are essential tools in the study of biological processes and are employed to monitor changes in gene expression and protein levels. Luciferases are reporter proteins that enable rapid and highly sensitive detection with an outstanding dynamic range. Here we evaluated the usefulness of the 19?kDa luciferase NanoLuc (Nluc), derived from the deep sea shrimp Oplophorus gracilirostris, as a reporter protein in yeast. Cassettes with codon-optimized genes expressing yeast Nluc (yNluc) or its destabilized derivative yNlucPEST have been assembled in the context of the dominant drug resistance marker kanMX. The reporter proteins do not impair the growth of yeast cells and exhibit half-lives of 40 and 5?min, respectively. The commercial substrate Nano-Glo® is compatible with detection of yNluc bioluminescence in?

SUBMITTER: Masser AE

PROVIDER: S-EPMC5069653 | biostudies-literature | 2016 May

REPOSITORIES: biostudies-literature

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Luciferase NanoLuc as a reporter for gene expression and protein levels in Saccharomyces cerevisiae.

Masser Anna E AE Kandasamy Ganapathi G Kaimal Jayasankar Mohanakrishnan JM Andréasson Claes C

Yeast (Chichester, England) 20160317 5

Reporter proteins are essential tools in the study of biological processes and are employed to monitor changes in gene expression and protein levels. Luciferases are reporter proteins that enable rapid and highly sensitive detection with an outstanding dynamic range. Here we evaluated the usefulness of the 19 kDa luciferase NanoLuc (Nluc), derived from the deep sea shrimp Oplophorus gracilirostris, as a reporter protein in yeast. Cassettes with codon-optimized genes expressing yeast Nluc (yNluc) ...[more]

PMID: 26860732

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