Gene Expression Analysis by Arylsulfatase Assays in the Green Alga Chlamydomonas reinhardtii

Methods Mol Biol. 2018:1755:149-161. doi: 10.1007/978-1-4939-7724-6_11.

Abstract

Chlamydomonas reinhardtii, a single-celled green alga, is a powerful microbial experimental system for understanding gene function. As a consequence of a high-quality genome sequence, community-wide efforts for gene model refinement and annotation, resources for strain collections and robust molecular techniques, research with this organism has significantly expanded in the past few decades. In two companion chapters, we outline colorimetric and fluorescence-based methodologies for genetic reporter systems in Chlamydomonas, which can be used to investigate and delineate gene expression and regulatory mechanisms. Here, we describe protocols for arylsulfatase activity assays using ARS2, activity of which can be measured either quantitatively or qualitatively, and in low (individual sample) or high (96-well format) throughput.

Keywords: ARS2; Algae; Gene fusion; High-throughput assay; Reporter.

Publication types

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

MeSH terms

  • Algal Proteins / genetics*
  • Arylsulfatases / genetics*
  • Arylsulfatases / metabolism
  • Base Sequence
  • Biological Assay / instrumentation
  • Biological Assay / methods
  • Chlamydomonas reinhardtii / genetics*
  • Colorimetry / instrumentation
  • Colorimetry / methods
  • Electroporation / instrumentation
  • Electroporation / methods
  • Enzyme Assays / instrumentation
  • Enzyme Assays / methods*
  • Escherichia coli
  • Gene Expression Regulation
  • Genes, Reporter / genetics*
  • Spectrometry, Fluorescence / instrumentation
  • Spectrometry, Fluorescence / methods
  • Transformation, Bacterial

Substances

  • Algal Proteins
  • Arylsulfatases