Plant Molecular Biology

, Volume 55, Issue 6, pp 869–881

Monitoring dynamic expression of nuclear genes in Chlamydomonas reinhardtii by using a synthetic luciferase reporter gene

  • Markus Fuhrmann
  • Amparo Hausherr
  • Lars Ferbitz
  • Thomas Schödl
  • Markus Heitzer
  • Peter Hegemann
Article

DOI: 10.1007/s11103-005-2150-1

Cite this article as:
Fuhrmann, M., Hausherr, A., Ferbitz, L. et al. Plant Mol Biol (2004) 55: 869. doi:10.1007/s11103-005-2150-1

Abstract

For monitoring the expression profile of selected nuclear genes in Chlamydomonas reinhardtii in response to altered environmental parameters or during cell cycle, in the past many RNA or protein samples had to be taken and analyzed by RNA hybridization or protein immunoblotting. Here we report the synthesis of a gene that codes for the luciferase of Renilla reniformis (RLuc) and is adapted to the nuclear codon usage of C. reinhardtii. This crluc gene was expressed alone or as a fusion to the zeocin resistance gene ble under control of different promoter variants. Luciferase activity was monitored in living cells, increased with the promoter strength and paralleled the amount of expressed protein. Under control of the Lhcb-1 promoter the Luc-activity in synchronized cultures was dependent on the dark-light cycle. Additionally, crluc was placed under control of the Chop-2 promoter and activity was measured under different light conditions. Chop-2 promoter activity was found to be most pronouced under low-light and dark conditions, further supporting that channelrhodopsin-2 is most active in dark-adapted cells. We conclude that crluc is a reliable tool for convenient monitoring of nuclear gene expression in C. reinhardtii.

Key words

bioluminescence channelrhodopsin-2 Chop-2 gene expression synthetic gene 

Abbreviations

Chop-1/2

channelopsin-1/2

ChR1/2

channelrhodopsin-1/2

HSM

high salt medium

Lhc

light-harvesting complex

Luc

luciferase

mAb

monoclonal antibody

RbcS2

small subunit of ribulosebisphosphate-carboxylase/oxygenase

RCF

relative codon frequency

RLU

relative luminescence units

TAP

Tris/acetate/phosphate medium

UTR

untranslated region

wt

wild type

Copyright information

© Kluwer Academic Publishers 2004

Authors and Affiliations

  • Markus Fuhrmann
    • 1
    • 2
  • Amparo Hausherr
    • 1
  • Lars Ferbitz
    • 1
  • Thomas Schödl
    • 1
  • Markus Heitzer
    • 2
  • Peter Hegemann
    • 1
  1. 1.Institut für Biochemie IUniversität RegensburgRegensburgGermany
  2. 2.Center of excellence for fluorescent bioanalysis, Biotechnology DivisionUniversität RegensburgRegensburgGermany
  3. 3.MünchenGermany
  4. 4.Institute for Molecular Biology and Biophysics ETHZZuerichSwitzerland
  5. 5.GeneArt GmbHRegensburgGermany

Personalised recommendations