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the complete nucleotide sequence of tobacco necrosis virus strain d.the complete sequence of the rna genome of tobacco necrosis virus strain d (tnv-d) consisting of 3759 nucleotides has been determined. the positive strand contains five open reading frames (orfs). the 5'-proximal orf encodes a 22k protein terminating with an amber codon which may be read through to produce a 82k protein (p82). two small centrally located orfs each encode two out-of-frame 7k proteins (p7a and p7b). the 3'-proximal orf encodes the 29k coat protein (cp), the n terminus of which has ...19911856691
molecular cloning and sequencing of the coat protein gene of a nebraskan isolate of tobacco necrosis virus: the deduced coat protein sequence has only moderate homology with those of strain a and strain d.a partial nucleotide sequence spanning the coat protein (cp) gene of a nebraskan isolate of tobacco necrosis virus (tnv-ne) has been determined. the sequence contains at least four open reading frames (orfs). the 5'-terminal orf encodes a protein that has 86% and 38% homology with the polymerases of strains a (tnv-a) and d (tnv-d), respectively. the second and third orfs probably encode 10.7 kda and 6.2 kda proteins (p 10.7 and p 6.2). these are respectively 90% and 96% amino acid homologous enc ...19938379851
specificity of satellite activation by tobacco necrosis virus correlates with nucleic acid hybridization pattern between helper virus isolates.tobacco necrosis virus (tnv) comprises over 20 different isolates which are usually classified on the basis of serological cross-reactivity of their virus particles or specific activation of satellite virus strains (stnv-1, -2, and -c). we have studied the relationships between five tnv isolates, tnv-a, -g, -cn, -d, and -ac36 which exhibit considerable differences in symptom formation on phaseolus vulgaris. it is shown that, like tnv-a, tnv-g and -cn support the multiplication of stnv-1 and -2. ...19938460499
nucleotide sequence of the genome of a citrus isolate of olive latent virus 1.the 3699 nt genome of olive latent virus 1 (olv-1), described years ago from southern italy as a putative sobemovirus, was completely sequenced. olv-1 genomic rna was not polyadenylated and had a structure virtually identical to that of species of the necrovirus rather than the sobemovirus genus. five open reading frames (orfs) were identified, of which the 5'-proximal encoded a 23k protein and ended with an amber codon whose readthrough could yield a putative 82k product. this polypeptide had e ...19968678829
complete nucleotide sequence of tobacco necrosis virus strain dh and genes required for rna replication and virus movement.the complete genome sequence of tobacco necrosis virus strain d (hungarian isolate, tnv-dh) was determined. the genome (3762 nt) has an organization identical to that reported for tnv-d. highly infectious synthetic transcripts from a full-length tnv-dh cdna clone were prepared, the first infectious necrovirus transcript reported. this clone was used for reverse genetic studies to map the viral genes required for replication and movement. protoplast inoculation with delta 22 and delta 82 mutants ...19979191913
the nucleotide sequence of satellite tobacco necrosis virus strain c and helper-assisted replication of wild-type and mutant clones of the virus.the complete nucleotide sequence of satellite tobacco necrosis virus strain c (stnv-c) was determined. the genome has a similar overall organization to two stnv isolates studied previously but differs significantly from them in the secondary structure of the translated and untranslated regions (utrs). stnv-c rna is naturally uncapped and contains 1221 nt: 101 nt in the 5' utr, 606 nt in the capsid protein (cp) coding region and 514 nt in the 3' utr. using the known sequences of stnv-c and tobacc ...19989634099
isolation of tobacco necrosis virus from strawberry leaves in the czech republic.leaves of symptomless fragaria ananassa duch cv. cacanská raná were grafted onto fragaria vesca indicator clones. thirty-five of 72 grafted indicator plants developed leaf mottle symptoms. isometric virus-like particles were observed in purified preparations from symptomatic leaves of f. vesca. the latter were mechanically inoculated to herbaceous host plants. a virus was successfully purified from nicotiana occidentalis 37 b symptomatic plants by differential and sucrose density gradient centri ...199810358734
mutation analysis of cis-elements in the 3'- and 5'-untranslated regions of satellite tobacco necrosis virus strain c rna.the putative, 3'-terminal stem-loop structure in satellite tobacco necrosis virus strain c (stnv-c) rna constitutes an essential cis-acting structure for the promotion of negative-strand rna synthesis and a single-stranded tail is also important. the putative, 5'-terminal stem-loop structure in stnv-c rna is not essential for productive, plus-strand rna accumulation but is required for optimal accumulation. residues 2 and 3 are the minimal cis-acting sequences required for rna synthesis. the rna ...199910544131
preferential banding of secondary veins in strawberry is caused by mixed virus infection.leaves of fragaria ananassa duch. cv. redgauntlet with mottle and mild dwarf symptoms were grafted onto f. vesca indicator clones. the youngest leaves developed specific vein banding pattern located preferentially on secondary veins near the edge of the leaves. electron microscopy of ultrathin sections and negatively stained purified virus preparations from symptom-bearing strawberry leaves revealed presence of different-sized isometric virions. particles of about 50 nm and 23 nm in diameter wer ...199910825923
the complete nucleotide sequence of beet black scorch virus (bbsv), a new member of the genus necrovirus.the complete nucleotide sequence of beet black scorch virus (bbsv) was determined. the bbsv genome is composed of 3641 nucleotides and has similar organization with tobacco necrosis virus d of 61% nucleotide identity. the 5'-proximal open reading frame (orf) encodes a putative 23 kda protein and a 82 kda protein by reading-through of an amber termination codon. three small orfs located in the center of the genome may encode for a 4.2 kda protein and two 7 kda proteins. the 3'-proximal orf encode ...200212491108
a tobacco necrosis virus d isolate from olea europaea l.: viral characterization and coat protein sequence analysis.a virus isolated from olea europaea l. grown in portugal, was identified as a member of the species tobacco necrosis virus d (tnv-d, genus necrovirus, family tombusviridae), based on the molecular and serological properties of the purified virus particles. the genomic region encoding the coat protein (cp) of this isolate (named gp isolate) was amplified by rt-pcr and the cdna was cloned and sequenced. the cp gene encodes a predicted protein of 269 amino acids showing high identity (86.2%) to tnv ...200415168200
the complete genome sequence of a new necrovirus isolated from olea europaea l.the complete nucleotide sequence of a virus isolated from olea europaea l. (gp isolate), previously identified as an isolate of tobacco necrosis virus d (tnv-d) based on its coat protein sequence, was determined. the viral rna genome consists of 3683 nucleotides and contains five open reading frames. the putative rna-dependent rna polymerase shows 91.2% amino acid identity with that of an isolate of olive latent virus 1 (olv-1) and the coat protein reveals highest sequence identity with that of ...200415592885
complete genome sequence of a tobacco necrosis virus d isolate from olive trees. 200919506797
subgenomic mrna transcription in tobacco necrosis virus.tobacco necrosis virus-d (tnv-d), a positive-strand rna necrovirus in the family tombusviridae, transcribes two subgenomic (sg) mrnas during infections. we have investigated the strategy used by tnv-d in this process and uncovered evidence that it employs a premature termination (pt) mechanism for the transcription of its sg mrnas. structural and mutational analysis of the tnv-d genome identified local rna structures upstream from transcriptional initiation sites that functioned in the plus-stra ...201121803392
a degenerate pair of primers for simultaneous detection of four alpha- and betanecroviruses.the high infection levels due to olive latent virus 1 (olv-1), olive mild mosaic virus (ommv) (alphanecrovirus) and tobacco necrosis virus d (tnv-d) (betanecrovirus) in portuguese olive orchards prompted us to develop a rapid pcr-based assay for the simultaneous detection of these viruses aimed at the sanitary selection and marketing of plant material in compliance with european union regulations. a pair of degenerate oligonucleotide primers, parrdrp5' and parcoat3' was designed based on conserv ...201425116202
correction: genetic diversity of the coat protein of olive mild mosaic virus (ommv) and tobacco necrosis virus d (tnv-d) isolates and its structural implications. 201525955830
genetic diversity of the coat protein of olive mild mosaic virus (ommv) and tobacco necrosis virus d (tnv-d) isolates and its structural implications.the genetic variability among 13 isolates of olive mild mosaic virus (ommv) and of 11 isolates of tobacco necrosis virus d (tnv-d) recovered from olea europaea l. samples from various sites in portugal, was assessed through the analysis of the coat protein (cp) gene sequences. this gene was amplified through reverse transcriptase polymerase chain reaction (rt-pcr), cloned, and 5 clone sequences of each virus isolate, were analysed and compared, including sequences from ommv and tnv-d isolates or ...201425350108
untranslated regions of diverse plant viral rnas vary greatly in translation enhancement efficiency.whole plants or plant cell cultures can serve as low cost bioreactors to produce massive amounts of a specific protein for pharmacological or industrial use. to maximize protein expression, translation of mrna must be optimized. many plant viral rnas harbor extremely efficient translation enhancers. however, few of these different translation elements have been compared side-by-side. thus, it is unclear which are the most efficient translation enhancers. here, we compare the effects of untransla ...201222559081
atypical rna elements modulate translational readthrough in tobacco necrosis virus d.tobacco necrosis virus, strain d (tnv-d), is a positive-strand rna virus in the genus betanecrovirus and family tombusviridae the production of its rna-dependent rna polymerase, p82, is achieved by translational readthrough. this process is stimulated by an rna structure that is positioned immediately downstream of the recoding site, termed the readthrough stem-loop (rtsl), and a sequence in the 3' untranslated region of the tnv-d genome, called the distal readthrough element (drte). notably, a ...201728148800
protein expression strategies in tobacco necrosis virus-d.tobacco necrosis virus (tnv-d) has a plus-strand rna genome that is neither 5' capped nor 3' poly-adenylated. instead, it utilizes a 3' cap-independent translational enhancer (3'cite) located in its 3' untranslated region (utr) for translation of its proteins. we have examined the protein expression strategies used by tnv-d and our results indicate that: (i) a base pairing interaction between conserved acca and uggu motifs in the genomic 5'utr and 3'cite, respectively, is not required for effici ...201526402375
translational readthrough in tobacco necrosis virus-d.the plus-strand rna genome of tobacco necrosis virus-d (tnv-d) expresses its polymerase via translational readthrough. the rna signals involved in this readthrough process were characterized in vitro using a wheat germ extract translation system and in vivo via protoplast infections. the results indicate that (i) tnv-d requires a long-range rna-rna interaction between an extended stem-loop (sl) structure proximal to the readthrough site and a sequence in the 3'-untranslated region of its genome; ...201424503089
investigations on the tobacco necrosis virus d p60 replicase protein.tobacco necrosis virus d (tnv-d), in the genus betanecrovirus (family tombusviridae), possesses a single-stranded, positive-sense rna genome containing six open reading frames (orfs). two 5'-proximal orfs (1 and 2) encode overlapping polypeptides of 22 and 82 kda (p22 and p82, respectively) which are both required for replication. the p22 auxiliary protein contains no replication motifs but the c-terminal region, downstream of a leaky stop codon, encodes a 60 kda polypeptide (p60) which contains ...201324278346
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