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The 5′-untranslated leader sequence of potato virus X RNA enhances the expression of a heterologous gene in vivo

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Summary

The 5′ untranslated leader of potato virus X (PVX) RNA is shown when contiguous to the coding sequence, to enhance the expression of the neomycin phosphotransferase II gene (NPTII) in Nicotiana tabacunz protoplasts in vivo. The level of transient expression of the NPTII gene in protoplasts provided by a plasmid containing the coding sequence of the NPTII gene under the control of 35S cauliflower mosaic virus (CaMV) promoter and terminator elements served as the baseline control. Insertion of the viral 5′ untranslated leader sequence upstream of the NPTII ATG codnn increased the level of expression 4-fold. An 83 nucleotide (nt) leader sequence (lacking only one nucleotide of the complete PVX leader) and a truncated version with a 28 deletion at the 3′ end both had similar enhancing abilities. The 28 nt CA region of the PVX leader alone had no enhancement properties.

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References

  • Browning KS, Lax SR, Humphreys J, Ravel JM (1988) Evidence that the 5′ untranslated leader of mRNA affects the requirement for wheat germ initiation factors 4A, 4F and 4G. J Biol Chem 263:9630–9634

    Google Scholar 

  • Gallie DR, Sleat DE, Watts JW, Turner PC, Wilson TMA (1987) The 5′ leader sequence of tobacco mosaic virus RNA enhances the expression of foreign gene transcripts in vitro and in vivo. Nucleic Acids Res 15:3257–3273

    Google Scholar 

  • Gallie DR, Sleat DE, Watts JW, Turner PC, Wilson TMA (1988a) Mutational analysis of tobacco mosaic virus 5′-leader for altered ability to enhance translation. Nucleic Acids Res 16:883–893

    Google Scholar 

  • Gallic DR, Walbot V, Hershey JWB (1988b) The ribosomal fraction mediates the translational enhancement associated with the 5′-leader of tobacco mosaic virus. Nucleic Acids Res. 16:8675–8694

    Google Scholar 

  • Herrera-Estrella L, Depicker A, Van Montagu M, Schell J (1983) Expression of chimeric genes transferred into plants cells using a Ti-plasmid derived vector. Nature (London) 303:209–213

    Google Scholar 

  • Jobling SA, Gehrke L (1987) Enhanced translation of chimeric messenger RNAs containing a plant viral untranslated leader sequence. Nature 325:622–625

    Google Scholar 

  • Kozak M (1986) Point mutations define a sequence flanking the AUG initiator codon that modulates translation by eukaryotic ribosomes. Cell 44:283–292

    Google Scholar 

  • Negrutiu I, Shillito R, Potrykus I, Biasini G, Sala F (1987) Hybrid genes in analysis of transformation conditions. Plant Mol Biol 8:363–373

    Google Scholar 

  • Paszkowski J, Shillito R, Saul M, Mandak V, Hohn T, Hohn B, Potrykus I (1984) Direct gene transfer to plants. EMBO J 3:2717–2722

    Google Scholar 

  • Reiss B, Sprengel R, Schaller H (1984a) Protein fusions with the kanamycin resistance gene from transposon Tn5. EMBO J 3:3317–3322

    Google Scholar 

  • Reiss B, Sprengel R, Will H, Schaller H (1984b) A new sensitive method for qualitative and quantitative assay of neomycin phosphotransferase in crude cell extracts. Gene 30:211–218

    Google Scholar 

  • Skryabin KG, Kraev AS, Morozov SYu, Rozanov MN, Chernov BK, Lukasheva LI, Atabekov IG (1988) The nucleotide sequence of potato virus X RNA. Nucleic Acids Res 16:10929–10930

    Google Scholar 

  • Smirnov OYu, Tashpulatov AS, Zakharyev VM, Skryabin KG (1987) Construction of chimeric marker genes NOS-NPT for plants. Bioorg Khimiya 13:1142–1145

    Google Scholar 

  • Sleat DE, Gallie DR, Jefferson RA, Bevan MW, Turner PV, Wilson TMA (1987) Characterization of 5′-leader sequence of tobacco mosaic virus RNA as a general enhancer of translation in vitro. Gene 217:217–225

    Google Scholar 

  • Sleat DE, Hull R, Turner PC, Wilson TMA (1988) Studies on mechanism of translation enhancement by the 5′-leader sequence of tobacco mosaic virus RNA. Eur J Biochem 175:75–86

    Google Scholar 

  • Töpfer R, Matzeit V, Gronenborn B, Schell J, Steinbiss H-H (1987) A set of plant vectors for transcriptional and translational fusions. Nucleic Acids Res 15:5890

    Google Scholar 

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Authors and Affiliations

  1. Centre of Bioengineering, Academy of Sciences of Russia, Vavilov Street 32, 117984, Moscow, Russia

    Mikhail M. Pooggin & Konstantin G. Skryabin

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  1. Mikhail M. Pooggin
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  2. Konstantin G. Skryabin
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Communicated by G.P. Georgiev

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Pooggin, M.M., Skryabin, K.G. The 5′-untranslated leader sequence of potato virus X RNA enhances the expression of a heterologous gene in vivo. Molec. Gen. Genet. 234, 329–331 (1992). https://doi.org/10.1007/BF00283854

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  • DOI: https://doi.org/10.1007/BF00283854

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