In Vivo Analyses of Viral RNA Translation

  • William R. Staplin
  • W. Allen Miller
Part of the Methods in Molecular Biology™ book series (MIMB, volume 451)


Positive-strand RNA viruses often use noncanonical strategies to usurp the host translational machinery for their own benefit. These strategies have been analyzed using transient expression assays in the absence of replication, with reporter genes replacing viral genes. A sensitive and convenient reporter assay is the dual luciferase system using Renilla (Renilla reniformis) and firefly (Photinus pyralis) reporter genes. Use of recombinant viral constructs containing the reporter luciferase gene allows us to discern whether a particular RNA sequence or secondary structure elicits an effect on initiation of translation or recoding. This chapter describes a standard luciferase protocol that can be molded to fit any viral sequence, in order to detect cis-acting regulatory elements in viral RNA.


Cap-independent translation Electroporation Oat protoplast Plant cell suspension culture Recoding Ribosomal frameshift 


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Copyright information

© Humana Press, a part of Springer Science + Business Media, LLC 2008

Authors and Affiliations

  • William R. Staplin
    • 1
  • W. Allen Miller
    • 1
  1. 1.Molecular Cellular and Developmental Biology, Department of Plant PathologyIowa State UniversityAmesUSA

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