Abstract
The genome of the tobacco mosaic virus (TMV) strain V-69 was sequenced by us earlier. The comparison of its nucleotide sequence with those of well-known tobamovirus strains (using published data) demonstrated considerable homology between strain V-69 and tomato strain L of TMV. In this study, full-length cDNA copy of TMV strain V-69 genomic RNA has been cloned in vector pBS(+) under the control of the T7 promoter and with an SmaI site at the 3’-terminus. The in vitro transcription of this construction with the use of phage T7 RNA polymerase in the presence of a cap analog (m 7 GpppG) has yielded an infectious RNA. This RNA has induced the same symptoms as those caused by TMV strain V-69 in infected indicator plants. Three point mutations inactivating one of two HindIII sites have been introduced into the cDNA. The absence of the HindIII site at position 1449 of the viral nucleotide sequence and the corresponding mutations serve as markers of the construction obtained. The full-length cDNA copy of the TMV strain V-69 RNA can be used both as a research tool and for constructing a vector for foreign gene expression.
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REFERENCES
H.R.B. Pelham (1978) ArticleTitleLeaky UAG Termination Codon in Tobacco Mosaic Virus RNA Nature 272 469–471
J.M. Skuzeski L.M. Nichols R.F. Gesteland J.F. Atkins (1991) ArticleTitleThe Signal for a Leaky UAG Stop Codon in Several Plant Viruses Includes the Two Downstream Codons J. Mol. Biol. 218 IssueID2 365–373
T.R. Hunter T. Hunt I. Knowland D. Zimmern (1976) ArticleTitleMessenger RNA for the Coat Protein of Tobacco Mosaic Virus Nature 260 759–764
A. Siegel V. Hari K. Kolacz (1978) ArticleTitleThe Effects of Tobacco Mosaic Virus Infection on Host and Virus-Specific Protein Synthesis in Protoplasts Virology 85 494–503
V.Z. Grdzelishvili S.N. Chapman W.O. Dawson D.J. Lewandowski (2000) ArticleTitleMapping of the Tobacco Mosaic Virus Movement Protein and Coat Protein Subgenomic RNA Promoters in Vivo Virology 275 177–192
N.R. Beachy M. Heinlein (2000) ArticleTitleRole of P30 in Replication and Spread of TMV Traffic 1 540–544
W.O. Dawson (1992) ArticleTitleTobamovirus-Plant Interactions Virology 186 359–367
N. Banerjee J.Y. Wang M. Zaitlin (1995) ArticleTitleA Single Nucleotide Change in the Coat Protein Gene of Tobacco Mosaic Virus Is Involved in the Induction of Severe Chlorosis Virology 207 234–239
M.A. Sulzinski K.A. Gabaro P. Palukaitis M. Zaitlin (1985) ArticleTitleReplication of Tobacco Mosaic Virus: VII. Characterization of a Third Subgenomic TMV RNA Virology 145 132–140
K.S. Sukhov T.S. Pod’‘yapol’skaya L.I. Izvekova et al. (1982) ArticleTitleTobacco Mosaic Virus Vaccine Strain V-69: Construction, Properties, and Application Izv. Akad. Nauk SSSR, Ser. Biol. 1 113–125
A.N. Shiyan N.V. Mil’shina P.B. Snegireva V.A. Pukhal’skii (1994) ArticleTitlePrimary Structure Analysis of TMV Vaccine Strain V-69 Rus. J. Genet. 30 IssueID12 1406–1408
P.B. Snegireva A.N. Shiyan E.A. Istomina (1999) ArticleTitleComplete Nucleotide Sequence of the V-69 Tobacco Mosaic Virus Vaccine Strain Rus. J. Genet. 35 IssueID7 748–754
T. Ohno M. Aoyagi Y. Yamanashi et al. (1984) ArticleTitleNucleotide Sequence of the TMV (Tomato Strain) Genome and Comparison with the Common Strain Genome J. Biochem. 96 1915–1923
J. Donson C.M. Kearney M.E. Hilf O.W. Dawson (1991) ArticleTitleSystemic Expression of a Bacterial Gene by a Tobacco Mosaic Virus-Based Vector Proc. Natl. Acad. Sci. USA 88 7204–7208
H. Hamamoto Y. Sugiyama N. Nakagawa et al. (1993) ArticleTitleA New Tobacco Mosaic Virus Vector and Its Use for the Systemic Production of Angiotensin I-Converting Enzyme Inhibitor in Transgenic Tobacco and Tomato Biotechnology 11 930–932
S. Shivprasad G.P. Pogue D.J. Lewandowski et al. (1999) ArticleTitleHeterologous Sequences Greatly Affect Foreign Gene Expression in Tobacco Mosaic Virus-Based Vectors Virology 255 312–323
S.G. Gribanov A.V. Shcherbakov N.A. Perevozchikova A.A. Gusev (1996) ArticleTitleUse of Aerosil A-300 and Filters GF/F (GF/S) for Purifying DNA Fragments, plasmid DNA, and RNA Biokhimiya 61 IssueID6 1064–1070
J. Sambrook E.F. Fritsch T. Maniatis (1989) Molecular Cloning: A Laboratory Manual New York: Cold Spring Harbor Lab. Cold Spring Harbor
M.J. Carter I.D. Milton (1993) ArticleTitleAn Inexpensive and Simple Method for DNA Purification on Silica Particles Nucleic Acids Res. 21 1044
H.J. None H. Nojima H. Okayma (1990) ArticleTitleHigh Efficiency Transformation of E. coli with Plasmids Gene 96 23–28 Occurrence Handle1:CAS:528:DyaK3MXns1aqug%3D%3D Occurrence Handle2265755
A.B. Vartapityan (1991) ArticleTitleThe Polymerase Chain Reaction Mol. Biol. 25 IssueID4 926–936
I.T. Atabekov (1981) Praktikum po obshchei virusologii (Laboratory Works on General Virology) Mosk. Gos. Univ. Moscow 60–73
A.A. Gusev (1997) ArticleTitleA Simple Method of RNA Isolation and Purification Bioorg. Khim. 23 IssueID9 763–765
M. Nishiguchi S. Kikuchi Y. Kiho et al. (1985) ArticleTitleMolecular Basis of Plant Viral Virulence: The Complete Nucleotide Sequence of an Attenuated Strain of Tobacco Mosaic Virus Nucleic Acids Res. 13 IssueID15 5585–5590
C.A. Holt R.A.J. Hodgson F.A. Coker et al. (1990) ArticleTitleCharacterization of the Masked Strain of Tobacco Mosaic Virus: Identification of the Region Responsible for Symptom Attenuation by Analysis of an Infectious cDNA Clone Mol. Plant-Microbe Interact. 3 IssueID6 417–423
D.J. Lewandowski W.O. Dawson (1993) ArticleTitleA Single Amino Acid Change in Tobacco Mosaic Virus Replicase Prevents Symptom Production Mol. Plant-Microbe Interact. 6 157–160
S. Asurmendi R.H. Berg J.C. Koo R.N. Beachy (2004) ArticleTitleCoat Protein Regulates Formation of Replication Complexes during Tobacco Mosaic Virus Infection Proc. Natl. Acad. Sci. USA 101 IssueID5 1415–1420
P. Sameer S.P. Goregaoker J.N. Culver (2003) ArticleTitleOligomerization and Activity of the Helicase Domain of the Tobacco Mosaic Virus 126-and 183-Kilodalton Replicase Proteins J. Virol. 77 IssueID6 3549–3556
A. dos Reis Figueira S. Golem S.P. Goregaoker J.N. Culver (2002) ArticleTitleA Nuclear Localization Signal and a Membrane Association Domain Contribute to the Cellular Localization of the Tobacco Mosaic Virus 126-kDa Replicase Protein Virology 301 81–89
T.A. Osman K.W. Buck (2003) ArticleTitleIdentification of a Region of the Tobacco Mosaic Virus 126-and 183-Kilodalton Replication Proteins Which Binds Specifically to the Viral 3’-Terminal tRNA-Like Structure J Virol. 77 IssueID16 8669–8675
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Translated from Genetika, Vol. 40, No. 12, 2004, pp. 1637–1645.
Original Russian Text Copyright © 2004 by Istomina, Snegireva, Shiyan.
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Istomina, E.A., Snegireva, P.B. & Shiyan, A.N. Construction of a full-length cDNA of tobacco mosaic virus strain v-69 genome. Russ J Genet 40, 1356–1363 (2004). https://doi.org/10.1007/s11177-005-0005-0
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DOI: https://doi.org/10.1007/s11177-005-0005-0