Document Detail

Internal ribosome entry site of Rhopalosiphum padi virus is functional in mammalian cells and has cryptic promoter activity in baculovirus-infected Sf21 cells.
MedLine Citation:
PMID:  18664329     Owner:  NLM     Status:  MEDLINE    
AIM: To substantiate the in vitro translational studies of a cross-kingdom, internal ribosome entry site (IRES), the 5 untranslated region of the Rhopalosiphum padi virus (RhPV), can function in mammalian cells and act as a shuttle IRES between insect cells and mammalian cells. METHODS: Cytomegalovirus (CMV) promoter-based bicistronic mammalian cell expression vectors, either in plasmids or baculovirus vectors, were generated. Plasmid transient transfection and baculovirus transduction assays were performed to test whether the RhPV IRES can mediate translation activity in versatile mammalian cell lines. RESULTS: Both plasmids and recombinant baculoviruses containing the CMV promoter and the RhPV IRES can mediate bicistronic gene expression in mammalian cells. However, in the CMV promoter containing recombinant baculovirus-infected insect Sf21 cells, only the second cistron gene expression was observed. Northern blot analysis and a promoterless assay demonstrated that the RhPV IRES exhibited cryptic promoter activity in baculovirus-infected insect cells. CONCLUSION: RhPV IRES can mediate gene expression in both insect cells and mammalian cells, and this characteristic of the RhPV IRES will facilitate the development of a bicistronic baculovirus gene therapy vectors.
Yi-jane Wu; Chao-yi Teng; Yu-jie Chen; Seng-chi Chen; Ying-ju Chen; Yi-ting Lin; Tzong-yuan Wu
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Acta pharmacologica Sinica     Volume:  29     ISSN:  1745-7254     ISO Abbreviation:  Acta Pharmacol. Sin.     Publication Date:  2008 Aug 
Date Detail:
Created Date:  2008-07-30     Completed Date:  2009-05-15     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  100956087     Medline TA:  Acta Pharmacol Sin     Country:  China    
Other Details:
Languages:  eng     Pagination:  965-74     Citation Subset:  IM    
Department of Bioscience Technology, Chung Yuan Christian University, Chungli 320, Taiwan, China.
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MeSH Terms
Baculoviridae / physiology*
Cell Line
Cytomegalovirus / genetics
Picornaviridae Infections / pathology,  virology*
Plasmids / genetics
Ribosomes / ultrastructure*
Transduction, Genetic

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