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

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 t...

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Published in:	Biotechnology and bioengineering Vol. 88; no. 6; pp. 730 - 739
Main Authors:	Shaloiko, L.A., Granovsky, I.E., Ivashina, T.V., Ksenzenko, V.N., Shirokov, V.A., Spirin, A.S.
Format:	Journal Article
Language:	English
Published:	Hoboken Wiley Subscription Services, Inc., A Wiley Company 20-12-2004
Subjects:	5' Untranslated Regions 5' Untranslated Regions - genetics 5′-UTR obelin mRNA leader cap-independent translation Cell-free system Cell-Free System - physiology cell-free translation systems Eukaryotic Cells - physiology Genetic Vectors Initiation factors Luminescent Proteins - biosynthesis Luminescent Proteins - genetics Obelia longissima Phages Plant Proteins - biosynthesis Plant Proteins - genetics Polyps Protein Biosynthesis - genetics Protein Engineering - methods Q1 Q2 Recombinant Proteins - biosynthesis RNA Cap Analogs - genetics sequestration of RNA-binding translation factors Transcription Translation translation inhibition by excess mRNA Triticum - embryology Triticum - genetics Triticum - metabolism Triticum aestivum Viruses - genetics Wheat germ
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Summary:	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. © 2004 Wiley Periodicals, Inc.
Bibliography:	istex:A2DC00143F29703FD5D4E0C52778CCD7B2FE7D02 INTAS Roche Molecular Biochemicals Program on Physico-Chemical Biology of the Russian Academy of Sciences - INTAS grant No. 99-00720 ArticleID:BIT20267 ark:/67375/WNG-F9CW3R13-D ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 ObjectType-Article-2 ObjectType-Feature-1
ISSN:	0006-3592 1097-0290
DOI:	10.1002/bit.20267