The 5'-leader sequence of tobacco mosaic virus RNA enhances the expression of foreign gene transcripts in vitro and in vivo

A 67-nucleotide portion of the non-coding, 5'-leader sequence of tobacco mosaic virus RNA [defined as omega' (Gr. omega prime)] has been shown to enhance the translation of contiguous foreign gene transcripts both in vitro and in vivo. Chemically-synthesized omega', containing conveni...

Full description

Saved in:
Bibliographic Details
Published in:Nucleic acids research 1987-04, Vol.15 (8), p.3257-3273
Main Authors: GALLIE, D. R, SLEAT, D. E, WATTS, J. W, TURNER, P. C, WILSON, T. M. A
Format: Article
Language:English
Subjects:
Animals
Base Sequence
Biological and medical sciences
Biotechnology
Cell-Free System
DNA, Recombinant - metabolism
Fundamental and applied biological sciences. Psychology
Gene Expression Regulation
Genetic engineering
Genetic technics
Genetic Vectors
Methods. Procedures. Technologies
Microinjections
Molecular cloning
Oocytes - metabolism
Protein Biosynthesis
Protoplasts - metabolism
Recombinant Fusion Proteins - biosynthesis
RNA Caps - physiology
RNA, Messenger - metabolism
RNA, Viral - physiology
Tobacco mosaic virus
Tobacco Mosaic Virus - genetics
Tobacco Mosaic Virus - physiology
Citations: Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:A 67-nucleotide portion of the non-coding, 5'-leader sequence of tobacco mosaic virus RNA [defined as omega' (Gr. omega prime)] has been shown to enhance the translation of contiguous foreign gene transcripts both in vitro and in vivo. Chemically-synthesized omega', containing convenient linker sequences, was inserted into derivatives of an in vitro transcription plasmid (pSP64) between the bacteriophage-SP6 promoter and sequences coding for either chloramphenicol acetyltransferase (CAT) or neomycin phosphotransferase (NPTII). Run-off in vitro transcripts, with or without a 5'-cap structure (G(5')ppp(5')G) and/or the omega' sequence, were tested in mRNA-dependent cell-free translation systems derived from rabbit reticulocyte lysate, wheat germ extract or Escherichia coli (MRE 600). In all cases, the presence of omega' increased the translational expression of both reporter genes, typically between 2- to 10-fold. Electroporation of isolated mesophyll protoplasts from Nicotiana tabacum cv. Xanthi, or microinjection of oocytes from Xenopus laevis, with SP6-transcripts containing the CAT-coding region confirmed and extended the value of omega' as a potential translational enhancer of gene expression in vivo.
ISSN:0305-1048
1362-4962
DOI:10.1093/nar/15.8.3257