Effective non-viral leader for cap-independent translation in a eukaryotic cell-free system.

Abstract

The 61 nt 5'-untranslated region (5'-UTR) of mRNA encoding for a light-emitting protein of hydroid polyp Obelia longissima, obelin, is shown to provide a high level of cap-independent translation of heterologous mRNAs in cell-free translation systems based on wheat germ extracts. The inhibition of translation typically observed when excess mRNA is present or produced in a eukaryotic system (the so-called self-inhibition phenomenon) is found abated with mRNA constructs carrying the obelin mRNA leader. The role of the sequestration of a limiting initiation factor, probably eIF4F, in the self-inhibition phenomenon and the possible independence of the obelin mRNA leader from eIF4F are discussed. We propose the obelin mRNA leader be used for effective cap-independent translation in eukaryotic cell-free systems, including combined transcription-translation systems with uncontrolled phage polymerase-catalyzed accumulation of mRNA.

• Authors: L A Shaloiko, I E Granovsky, T V Ivashina, V N Ksenzenko, V A Shirokov, A S Spirin
• Journal: Biotechnology and bioengineering (Biotechnol Bioeng) Vol. 88 Issue 6 Pg. 730-9 (Dec 20 2004) ISSN: 0006-3592 [Print] United States
• PMID: 15532099 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• 5' Untranslated Regions
• Luminescent Proteins
• Plant Proteins
• RNA Cap Analogs
• Recombinant Proteins
• obelin

Topics

• 5' Untranslated Regions (genetics)
• Cell-Free System (physiology)
• Eukaryotic Cells (physiology)
• Genetic Vectors
• Luminescent Proteins (biosynthesis, genetics)
• Plant Proteins (biosynthesis, genetics)
• Protein Biosynthesis (genetics)
• Protein Engineering (methods)
• RNA Cap Analogs (genetics)
• Recombinant Proteins (biosynthesis)
• Triticum (embryology, genetics, metabolism)
• Viruses (genetics)