Plant Molecular Biology Reporter

, Volume 21, Issue 3, pp 271–280 | Cite as

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

  • Christopher Ian Cazzonelli
  • Jeff Velten
Genetic Resources


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.

Key words

dual luciferase assay firefly intron luciferase Photinus pyralis Renilla reniformis reporter gene sea pansy 


CaMV 35S

cauliflower mosaic virus 35S RNA promoter










Photinus pyralis (firefly) luciferase


Photinus pyralis (firefly) intron-modified luciferase


luciferase reagent II (Promega)


magnesium chloride


Murashige and


ng, nanograms


nopaline synthase terminator


neomycin phosphotransferase II


pBluescript II (SK+)


stop and glo (Promega)


primary transformant


progeny of self-fertilized R0 plant


Renilla reniformis (sea pansy) luciferase


Renilla reniformis (sea pansy) intron-modified luciferase


relative light units


tris-borate EDTA




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Copyright information

© International Society for Plant Molecular Biology 2003

Authors and Affiliations

  1. 1.United States Department of Agriculture-Agricultural Research ServicesUSDA-ARSUSA

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