Document Detail


Functional characterization of the EMCV IRES in plants.
MedLine Citation:
PMID:  11123797     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The translation of eukaryotic messenger RNA is typically dependent upon the presence of an m7GpppN cap structure at the 5' end of the transcript. However, several animal viruses, including the Picorna viruses, have been shown to exhibit cap-independent translation through the presence of an internal ribosome entry site or IRES. This IRES-mediated cap-independent internal translation initiation has been exploited to generate bicistronic transcripts that function in animal cells. Recently IRES elements have also been identified in a small number of vertebrate, insect and yeast cellular messenger RNAs although no such sequences have been identified in endogenous plant genes and there are no reports of animal virus derived IRES activity in plant cells. Here we have constructed a bicistronic gene containing both green fluorescent protein and luciferase open-reading frames separated by the encephalomyocarditis IRES element under the control of the CaMV 35S promoter. Northern analysis reveals expression of the bicistronic transcript and in vivo imaging of GFP and luciferase activities demonstrates the functional presence of both proteins. Western blot analysis confirms the independent translation of both reporter proteins. These data suggest that insertion of the encephalomyocarditis virus (EMCV) IRES element between two open-reading frames of a plant bicistronic transcript can mediate translation of the second open-reading frame. This activity is more apparent in the leaves, than in the roots, of transgenic seedlings carrying the bicistronic reporter gene construct.
Authors:
P Urwin; L Yi; H Martin; H Atkinson; P M Gilmartin
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  The Plant journal : for cell and molecular biology     Volume:  24     ISSN:  0960-7412     ISO Abbreviation:  Plant J.     Publication Date:  2000 Dec 
Date Detail:
Created Date:  2001-01-02     Completed Date:  2001-03-08     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  9207397     Medline TA:  Plant J     Country:  ENGLAND    
Other Details:
Languages:  eng     Pagination:  583-9     Citation Subset:  IM    
Affiliation:
Centre for Plant Sciences, Leeds Institute for Plant Biotechnology and Agriculture, University of Leeds, Leeds LS2 9JT, UK.
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MeSH Terms
Descriptor/Qualifier:
DNA, Recombinant
Encephalomyocarditis virus / genetics*
Gene Expression Regulation, Plant
Green Fluorescent Proteins
Luciferases / genetics,  metabolism
Luminescent Proteins / genetics,  metabolism
Microscopy, Fluorescence
Open Reading Frames / genetics
Plants / genetics*
Plants, Genetically Modified
Plants, Toxic
RNA, Messenger / genetics,  metabolism
Recombinant Fusion Proteins / genetics,  metabolism
Regulatory Sequences, Nucleic Acid / genetics*
Tobacco / genetics
Transcription, Genetic
Chemical
Reg. No./Substance:
0/DNA, Recombinant; 0/Luminescent Proteins; 0/RNA, Messenger; 0/Recombinant Fusion Proteins; 147336-22-9/Green Fluorescent Proteins; EC 1.13.12.-/Luciferases

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