Renilla luciferase as a vital reporter for chloroplast gene expression in Chlamydomonas

I. Minko, S. P. Holloway, S. Nikaido, M. Carter, O. W. Odom, C. H. Johnson, D. L. Herrin

Research output: Contribution to journalArticle

49 Scopus citations

Abstract

The use of luciferases as reporters of gene expression in living cells has been extended to the chloroplast genome. We show that the luciferase from the soft coral Renilla reniformis (Rluc) can be successfully expressed in the chloroplast of Chlamydomonas reinhardtii. Expression of the rluc cDNA was driven by the promoter and 5' untranslated regions of the atpA gene. Western analysis with an anti-Rluc antibody detected a single polypeptide of 38 kDa in the luminescent cells. This is 3 kDa larger than native Rluc, and suggests that translation of the chimeric mRNA begins at the atpA start codon, 29 codons upstream from the rluc start site. We also show that the luminescence of the transformants was sufficient to enable imaging of colonies using a cooled CCD camera.

Original languageEnglish (US)
Pages (from-to)421-425
Number of pages5
JournalMolecular and General Genetics
Volume262
Issue number3
DOIs
StatePublished - Nov 25 1999

Keywords

  • Bioluminescence
  • Chlamydomonas
  • Chloroplast transformation
  • Chloroplast translation
  • Vital reporter

ASJC Scopus subject areas

  • Genetics

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    Minko, I., Holloway, S. P., Nikaido, S., Carter, M., Odom, O. W., Johnson, C. H., & Herrin, D. L. (1999). Renilla luciferase as a vital reporter for chloroplast gene expression in Chlamydomonas. Molecular and General Genetics, 262(3), 421-425. https://doi.org/10.1007/s004380051101