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Construction and testing of an intron-containing luciferase reporter gene fromRenilla reniformis

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We describe a newRenilla reniformis luciferase reporter gene,RiLUC, which was designed to allow detection of luciferase activity in studies involvingAgrobacterium-based transient expression studies. TheRLUC gene was altered to contain a modified intron from the castor bean catalase gene while maintaining consensus eukaryotic splicing sites recognized by the plant spliceosome.RLUC andRiLUC reporter genes were fused to the synthetic plant SUPER promoter. Luciferase activity within agrobacteria containing the SUPER-RLUC construct increased during growth in culture. In contrast, agrobacteria harboring the SUPER-RiLUC gene fusion showed no detectable luciferase activity. Agrobacteria containing these gene fusions were cotransformed with a compatible normalization plasmid containing a cauliflower mosaic virus 35S promoter (CaMV) joined to the firefly luciferase coding region (FiLUC) and infused into tobacco leaf tissues through stomatal openings. The kinetics of luciferase production from theRLUC orRiLUC reporters were consistent, with expression of theRiLUC gene being limited to transiently transformed plant cells.RiLUC activity from the reporter gene fusions was measured transiently and within stably transformed tobacco leaf tissues. Analysis of stably transformed tobacco plants harboring either reporter gene fusion showed that the intron altered neither the levels of luciferase activity nor tissue-specific expression patterns driven by the SUPER promoter. These results demonstrate that theRiLUC reporter gene can be used to monitor luciferase expression in transient and stable transformation experiments without interference from contaminating agrobacteria.

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CaMV 35S:

cauliflower mosaic virus 35S RNA promoter










Photinus pyralis (firefly) luciferase


Photinus pyralis (firefly) intron-modified luciferase


luciferase reagent II (Promega)

MgCl2 :

magnesium chloride


Murashige and


ng, nanograms

Nost :

nopaline synthase terminator


neomycin phosphotransferase II


pBluescript II (SK+)


stop and glo (Promega)

R0 :

primary transformant

R1 :

progeny of self-fertilized R0 plant


Renilla reniformis (sea pansy) luciferase


Renilla reniformis (sea pansy) intron-modified luciferase


relative light units


tris-borate EDTA




  • Becker D, Kemper E, Schell J, and Masterson R (1992) New plant binary vectors with selectable markers located proximal to the left T-DNA border. Plant Mol Biol 20(6): 1195–1197.

    Article  PubMed  CAS  Google Scholar 

  • Benfey P and Chua N (1990) The cauliflower mosaic virus 35s promoter: combinatorial regulation of transcription in plants. Sci 250(16): 959–966.

    Article  CAS  Google Scholar 

  • Bourdon V, Harvey A, and Lonsdale D (2001) Introns and their positions affect the translational activity of mRNA in plant cells. EMBO Rep 21(51): 394–398.

    Google Scholar 

  • Hajdukiewicz P, Svab Z, and Maliga P (1994) The small, versatile pPZP family ofAgrobacterium binary vectors for plant transformation. Plant Mol Biol 25: 989–994.

    Article  PubMed  CAS  Google Scholar 

  • Horsch R, Fry J, Hoffman N, Neidermeyer J, Rogers S, and Fraley R (1988) Plant Molecular Biology Manual: Leaf Disc Transformation, pp 1–9. Kluwer Academic Publishers, Belgium.

    Google Scholar 

  • Jefferson R, Kavanagh T, and Bevan M (1987)GUS fusions: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J 6(13): 3901–3907.

    PubMed  CAS  Google Scholar 

  • Leffel S, Mabon S, and Stewart N (1997) Applications of green fluorescent protein in plants. BioTechniques 23: 912–918.

    PubMed  CAS  Google Scholar 

  • Mankin S, Allen G, and Thompson W (1997) Introduction of a plant intron into the luciferase gene ofPhotinus pyralis. Plant Mol Biol Rep 15(2): 186–196.

    Article  CAS  Google Scholar 

  • Mayerhofer R, Langridge W, Cormier M, and Szalay A (1995) Expression of recombinantRenilla luciferase in transgenic plants results in high levels of light emission. Plant J 7: 1031–1038.

    Article  CAS  Google Scholar 

  • Ni M, Cui D, Einstein J, Narasimhulu S, Vergara Q, and Gelvin S (1995) Strength and tissue specificity of chimeric promoters derived from the octopine and mannopine synthase genes. Plant J 7(4): 661–676.

    Article  CAS  Google Scholar 

  • Ohta S, Mita S, Hattori T, and Nakamura K (1990) Construction and expression in tobacco of a β-glucuronidase (GUS) reporter gene containing an intron within the coding sequence. Plant Cell Physiol 31(6): 805–814.

    CAS  Google Scholar 

  • Ow D, Wood K, DeLuca M, DeWet J, Helsinki D, and Howell S (1986) Transient and stable expression of the firefly luciferase gene in plant cells and transgenic plants. Sci 234 (856–859).

    Article  CAS  Google Scholar 

  • Sambrook J and Russell D (2001) Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory, New York.

    Google Scholar 

  • Shapiro M and Senapathy P (1987) RNA splice junctions of different classes of eukaryotes: sequence statistics and functional implications in gene expression. Nucl Acids Res 15(17): 7155–7174.

    Article  PubMed  CAS  Google Scholar 

  • Svab Z, Hajdukiewicz P, and Maliga P (1995) Generation of transgenic tobacco plants by cocultivation of leaf disks withAgrobacterium pPZP binary vectors. In: Methods in Plant Molecular Biology: A Laboratory Course Manual, pp 55–77. Cold Spring Harbor Laboratory Press, New York.

    Google Scholar 

  • Vancanneyt G, Schmidt R, O'Connor-Sanchez A, Willmitzer L, and Rocha-Sosa M (1990) Construction of an intron-containing marker gene: splicing of the intron in transgenic plants and its use in monitoring early events inAgrobacterium-mediated plant transformation. Mol Gen Genet 220(2): 245–250.

    Article  PubMed  CAS  Google Scholar 

  • Walkerpeach C and Velten J (1994)Agrobacterium-mediated gene transfer to plant cells cointegrate and binary vector systems. In: Plant Molecular Biology Manual. pp 1–19 S. Gelvin and R. Schilperoort, Kluwer Academic Publishers, Belgium.

    Google Scholar 

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Correspondence to Jeff Velten.

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Cazzonelli, C.I., Velten, J. Construction and testing of an intron-containing luciferase reporter gene fromRenilla reniformis . Plant Mol Biol Rep 21, 271–280 (2003).

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