Molecular Cloning of Potato Leaf Roll Virus Complementary DNA

Smith, O. P.; Harris, Kerry F.; Toler, R. W.; Summers, Max D.

doi:10.1094/phyto-78-1060

Cited by 15 publications

(9 citation statements)

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“…Of the 28 sites described by Prill et al, the EcoRI site nearest the 5' end and the XhoI site nearest the 3' end are not in the sequence in Fig. 1 and eight sites in our sequence are not in the map shown by Prill et al All the 20 sites described by Smith et al (1988) are in the sequence shown in Fig. 1, but their positions are all about 100 nucleotides nearer the 5' end than they are in Fig.…”

Section: Nucleotide Sequencementioning

confidence: 43%

“…1 with the restriction endonuclease maps derived by Prill et al (1988) and Smith et al (1988) shows that most of the restriction sites are common to all three sequences, suggesting that there is little sequence difference between the Scottish, German and American isolates studied. Of the 28 sites described by Prill et al, the EcoRI site nearest the 5' end and the XhoI site nearest the 3' end are not in the sequence in Fig.…”

Section: Nucleotide Sequencementioning

confidence: 82%

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Nucleotide Sequence of Potato Leafroll Luteovirus RNA

Mayo¹,

Robinson²,

Jolly³

et al. 1989

Journal of General Virology

161

138

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SUMMARYA sequence of 5987 nucleotides is reported for the RNA of potato leafroll luteovirus (PLRV). The sequence contains six large open reading flames, and non-coding regions of 174 nucleotides at the 5' end, 141 nucleotides at the 3' end and 197 nucleotides between two large blocks of coding sequences. The 5' coding region encodes two polypeptides of 28 000 (28K) and 70K which overlap in different reading frames and circumstantial evidence suggests that the third open reading frame in the 5' block is translated by frameshift readthrough near the end of the 70K polypeptide to give a l18K polypeptide. The C-terminal part of the l18K protein contains the consensus sequence for RNA-dependent RNA polymerases. In vitro translation of PLRV RNA resulted in the synthesis mainly of 28K and 70K polypeptides and the largest product made was about 125K; these sizes are similar to those predicted for the translation products of the 5' block of coding sequence. The 3' block of coding sequence codes for three polypeptides: a 23K coat protein, a 17K polypeptide which is encoded in a different frame, and a 53K polypeptide which immediately follows the coat protein coding sequence, and is in the same reading frame. Circumstantial evidence suggests that the 53K polypeptide is translated by readthrough of the amber termination codon of the coat protein gene. The amino acid sequences encoded by the 3' block of coding sequence show many similarities with analogous polypeptides translated from the nucleotide sequences of RNA of barley yellow dwarf virus, PAV strain (BYDV) and,

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Section: Nucleotide Sequencementioning

confidence: 43%

Section: Nucleotide Sequencementioning

confidence: 82%

Nucleotide Sequence of Potato Leafroll Luteovirus RNA

Mayo¹,

Robinson²,

Jolly³

et al. 1989

Journal of General Virology

161

138

View full text Add to dashboard Cite

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“…Clone B2a and several other oligo(dT)-primed clones mapped to the same location suggesting that they were derived from the 3' end of PLRV RNA. Although some differences were observed, restriction analysis revealed overall similarities with the restriction sites of the PLRV isolates described by both Prill et al (1988) and Smith et al (1988).…”

supporting

confidence: 63%

Identification and Characterization of the Potato Leafroll Virus Putative Coat Protein Gene

Kawchuk

Martín

Rochon

et al. 1989

Journal of General Virology

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“…To determine the sequence cf the 3'end of Ob, RNA was polyadenylated (Smith et al, 1988) and first strand cDNAwas primed with an oligod(T) primer. Second strand synthesis was performed with a forward Ob primer (nucleotides 4891 to 4913), which was designed from the sequence derived from the PCR clones described above.…”

Section: Determining the Sequence Of The 3' End Of Obmentioning

confidence: 99%

Analysis of a tobacco mosaic virus strain capable of overcoming N gene-mediated resistance.

1993

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The genome of Ob, a tobamovirus that overcomes the N gene-mediated hypersensitive response (HR), was cloned as a cDNA, and its nucleotide sequence was determined. The genomic organization of Ob is similar to that of other tobamoviruses, consisting of 6506 nucleotides and containing at least four open reading frames. These open reading frames encode a 126-kD polypeptide with a 183-kD readthrough product, a 30.6-kD movement protein, and an 18-kD coat protein. A bacteriophage T7 promoter sequence was fused to the full-length cDNA clone to obtain infectious RNA transcripts. These transcripts, when inoculated onto tobacco plants, induced disease symptoms indistinguishable from plants inoculated with Ob viral RNA. To determine which viral factor is responsible for the resistance-breaking character of Ob, a recombinant virus was constructed in which the movement protein gene of tobacco mosaic virus was replaced with that of Ob. Cultivar Xanthi NN tobacco plants infected with this virus responded with an HR, indicating that the Ob movement protein alone does not act to overcome the N gene-mediated response. Following mutagenesis of the infectious Ob cDNA clone with hydroxylamine, populations of transcripts from the mutagenized DNA were inoculated onto Xanthi NN tobacco, and a variant that induced the HR was identified. The mutant was analyzed and found to contain a single nucleotide change in the 126-kD gene. Recreating the mutation in the Ob cDNA clone by site-directed mutagenesis resulted in a virus that caused symptoms identical to the chemically induced mutant.

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Molecular Cloning of Potato Leaf Roll Virus Complementary DNA

Cited by 15 publications

References 0 publications

Nucleotide Sequence of Potato Leafroll Luteovirus RNA

Nucleotide Sequence of Potato Leafroll Luteovirus RNA

Identification and Characterization of the Potato Leafroll Virus Putative Coat Protein Gene

Analysis of a tobacco mosaic virus strain capable of overcoming N gene-mediated resistance.

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