Using YFP as a Reporter of Gene Expression in the Green Alga Chlamydomonas reinhardtii

Methods Mol Biol. 2018:1755:135-148. doi: 10.1007/978-1-4939-7724-6_10.

Abstract

The unicellular green alga Chlamydomonas reinhardtii is a valuable experimental system in plant biology for studying metal homeostasis. Analyzing transcriptional regulation with promoter-fusion constructs in C. reinhardtii is a powerful method for connecting metal-responsive regulation with cis-regulatory elements, but overcoming expression-level variability between transformants and optimizing experimental conditions can be laborious. Here, we provide detailed protocols for the high-throughput cultivation of C. reinhardtii and assaying Venus fluorescence as a reporter for promoter activity. We also describe procedural considerations for relating metal supply to transcriptional activity.

Keywords: Algae; Copper; Iron; Yellow fluorescent protein; Zinc.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Bacterial Proteins / chemistry
  • Bacterial Proteins / genetics*
  • Biological Assay / instrumentation
  • Biological Assay / methods*
  • Chlamydomonas reinhardtii / genetics*
  • Gene Expression Regulation / genetics*
  • Gene Fusion / genetics
  • Gene Transfer Techniques / instrumentation
  • Genes, Reporter / genetics*
  • Luminescent Proteins / chemistry
  • Luminescent Proteins / genetics*
  • Response Elements / genetics

Substances

  • Bacterial Proteins
  • Luminescent Proteins
  • yellow fluorescent protein, Bacteria