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Use of animal viral internal ribosome entry site sequence makes multiple truncated transcripts without mediating polycistronic expression in rice

  • Harin Jung
  • Ju-Kon KimEmail author
  • Sun-Hwa HaEmail author
Biochemistry Article

Abstract

To establish a system that facilitates the coordinate expression of multiple genes in a crop biotechnology setting, we tested the internal ribosome entry site (IRES) sequence from encephalimyocarditis virus (an animal viral polycistronic sequence) in rice plants. Three reporter genes, green fluorescent protein (Gfp), β-glucuronidase (Gus) and luciferase (Luc), were linked to IRES under the control of the ubiquitin promoter, generating FIG (G f p-I RES-G us) and FIGIL (G f p-I RES-G us-I RES-L uc) constructs, respectively. We obtained a total 38 transgenic lines using these vectors (24 for FIC and 14 for FIGIL) and the transgene integration and copy numbers for each were determined by Southern blot analysis. By northern blot analysis using Gfp, Gus and Luc probes, both the FIG and FIGIL rice plants showed multiple truncated transcripts, including full-length recombinant genes. Western blot analyses using GFP and GUS antibodies revealed the accumulation of GFP but not GUS protein in both FIG and FIGIL rice leaf tissues. Subsequent GFP and GUS reporter expression patterns showed GFP fluorescence but not GUS-staining in both FIG and FIGIL transgenic rice callus tissues. These results suggest that the direct application of animal viral IRES sequences is not sufficient to induce polycistronic expression in rice plants due to the production of multiple truncated transcripts.

Key words

encephalimyocarditis virus internal ribosome entry site polycistronic rice 

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Copyright information

© Korean Society for Applied Biological Chemistry and Springer 2011

Authors and Affiliations

  1. 1.School of Biotechnology and Environmental EngineeringMyongji UniversityYonginRepublic of Korea
  2. 2.National Academy of Agricultural ScienceRural Development AdministrationSuwonRepublic of Korea

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