Establishment of a firefly luciferase reporter assay system in the unicellular red alga Cyanidioschyzon merolae

J Gen Appl Microbiol. 2021 Apr 16;67(1):42-46. doi: 10.2323/jgam.2020.02.003. Epub 2020 Sep 16.

Abstract

The firefly luciferase (Luc) reporter assay is a powerful tool used to analyze promoter activities in living cells. In this report, we established a firefly Luc reporter assay system in the unicellular model red alga Cyanidioschyzon merolae. A nitrite reductase (NIR) promoter-Luc fusion gene was integrated into the URA5.3 genomic region to construct the C. merolae NIR-Luc strain. Luc activities in the NIR-Luc strain were increased, correlating with the accumulation of endogenous NIR transcripts in response to nitrogen depletion. Luc activity was also significantly increased by the overexpression of the MYB1 gene, which encodes a transcription factor responsible for NIR promoter activation. Thus, our results demonstrate the utility of the Luc reporter system in C. merolae.

Keywords: Cyanidioschyzon merolae; luciferase; nitrogen; reporter assay; transcription factor.

MeSH terms

  • Gene Expression Regulation, Plant
  • Genes, Plant / genetics
  • Genes, Reporter*
  • Luciferases, Firefly / genetics*
  • Luciferases, Firefly / metabolism*
  • Nitrite Reductases
  • Plant Proteins / genetics
  • Plant Proteins / metabolism
  • Promoter Regions, Genetic
  • Rhodophyta / enzymology*
  • Rhodophyta / genetics*
  • Transcription Factors / genetics

Substances

  • Plant Proteins
  • Transcription Factors
  • Luciferases, Firefly
  • Nitrite Reductases