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  • 1
    Electronic Resource
    Electronic Resource
    Identification of different isolates of Cassava brown streak virus and development of a diagnostic test (2001)
    Oxford, UK : Blackwell Science Ltd
    Plant pathology 50 (2001), S. 0 
    Details
    ISSN: 1365-3059
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: An RT-PCR based detection method for Cassava brown streak virus (CBSV)-infected cassava has been developed. The RT-PCR detection method described includes RNA extraction methods for cassava leaves, a distinct primer set for the virus and RT-PCR conditions. The primers were designed to the virus coat protein gene and generate a virus-specific product of 231 bp from infected cassava. The test can detect the virus in the new growth of cassava sticks before any disease symptoms are visible. This test was used successfully with infected cassava from both Tanzania and Mozambique. Three isolates from Tanzania were found to exhibit different symptoms on the secondary host plants Nicotiana benthamiana and N. tabacum SR1. They have nucleotide sequence variation within the coat protein region of up to 8% and amino acid differences of up to 6%.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1365-3059.2001.00647.x
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  • 2
    Electronic Resource
    Electronic Resource
    Potato latent virus: a proposed new species in the genus Carlavirus (2002)
    Oxford, UK : Blackwell Science Ltd
    Plant pathology 51 (2002), S. 0 
    Details
    ISSN: 1365-3059
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: A previously undescribed carlavirus, potato latent virus (PotLV), was found infecting the potato cultivar Red La Soda imported from the USA. The particles were filamentous and slightly curved, with modal lengths of 530 and 670 nm. The 11 kDa protein encoded downstream from the coat protein contained a ‘zinc-finger’ motif characteristic of carlaviruses, and RT–PCR using a carlavirus-specific primer gave a PCR product of 857 bp. Antibodies produced to PotLV did not detect other carlaviruses when used in ELISA and the coat-protein nucleic acid sequence of PotLV showed 〈 67% similarity with the other carlaviruses tested. The closest similarity was with the Andean strain of potato virus S. Unusually for a carlavirus, PotLV systemically infected Nicotiana bigelovii, N. glutinosa, N. rustica, N. tabacum and Physalis floridana.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1365-3059.2002.00729.x
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  • 3
    Electronic Resource
    Electronic Resource
    Molecular characterization of the Cassava brown streak virus coat protein (2001)
    Oxford, UK : Blackwell Science Ltd
    Plant pathology 50 (2001), S. 0 
    Details
    ISSN: 1365-3059
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: A partial sequence of 1114 nucleotides of a virus from cassava brown streak diseased (CBSD) material was obtained. Alignment of the predicted amino acid sequence with those of other members of the Potyviridae showed closest identity with the coat protein of Sweet potato mild mottle virus (genus Ipomovirus). The predicted amino acid sequence has one open reading frame with a 3′ untranslated region of 144 nucleotides and a poly(A) tail. The expressed protein was shown to cross-react with an antiserum raised previously to a virus isolated from CBSD material. Evidence presented suggests that CBSD is caused by Cassava brown streak virus, a tentative member of the genus Ipomovirus, as this virus is consistently found associated with CBSD.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1365-3059.2001.00589.x
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  • 4
    Electronic Resource
    Electronic Resource
    Alternative tissue analysis method developed for organochlorine contaminants in aquatic organisms (1994)
    Springer
    Bulletin of environmental contamination and toxicology 53 (1994), S. 382-389 
    Details
    ISSN: 1432-0800
    Source: Springer Online Journal Archives 1860-2000
    Topics: Energy, Environment Protection, Nuclear Power Engineering , Medicine
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00197230
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  • 5
    Electronic Resource
    Electronic Resource
    In vivo characterisation of a translational enhancer upstream from the coat protein open reading frame of potato virus S (1994)
    Springer
    Archives of virology 137 (1994), S. 123-132 
    Details
    ISSN: 1432-8798
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The 101 nucleotide region upstream from the ATG of the potato virus S (PVS) coat protein gene was isolated and the effect of this region on the translation of a downstream open reading frame analysed in vivo. Translation was monitored using the reporter genes B-glucuronidase (GUS) and luciferase (LUC). Translational enhancement was assayed transiently using DNA microprojectile bombardment into both leaf and pollen tissue and also by polyethylene glycol mediated transfection of tobacco protoplasts. In both cases the presence of this region resulted in a 2–3 fold increase in translation when compared to reporter expression with synthetic leader and authentic plant leader constructs. Tobacco plants stabily transformed with this PVS 101 nucleotide region and downstream GUS gene gave 4 times the level of translation over synthetic leader GUS control plants.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01311178
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  • 6
    Electronic Resource
    Electronic Resource
    Analysis of a translational enhancer upstream from the coat protein open reading frame of potato virus S (1994)
    Springer
    Archives of virology 134 (1994), S. 321-333 
    Details
    ISSN: 1432-8798
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Evidence has suggested that the subgenomic RNA of the carlavirus potato virus S is an efficient message for the coat protein, even though evidence suggests it is uncapped at its 5′ terminus. We have investigated the effect of the upstream region of the coat protein gene of potato virus S on the level of reporter gene expression in vitro. The region of 101 nucleotides upstream of the coat protein, designated VTE (viral translational enhancer) was found to increase levels of translation in comparison to a synthetic leader when linked to the β-glucuronidase (GUS) reporter gene in vitro in rabbit reticulocyte and wheat germ lysate. VTE was also able to increase translation of the reporter gene luciferase (LUC) in vitro above the levels obtained for both a synthetic leader and a leader obtained from a plant gene isolated fromArabidopsis thaliana. The level of enhancement was evident with both capped and uncapped transcripts. When the VTE sequence was deleted to 20 nucleotides of the upstream region, thus removing the nucleotide block homologous among carlaviruses, the ability to enhance levels of translation was removed. In vitro translation studies indicated that the translational enhancement activity of VTE was at least partially cap independent. Translation of VTE linked to reporter genes in the presence of cap analogue was relatively unaffected whereas synthetic leader and a plant leader constructs were both more sensitive. In vitro competition analysis revealed that when short RNA transcripts representing the 101 nucleotides of VTE were added in trans to functional VTE leader LUC constructs there was a marked decrease in the level of translation when compared with a synthetic leader added in trans. These results suggest that the upstream region of the coat protein ORF of potato virus S promotes translation in a cap-independent manner that may involve the binding of proteins and/or ribosomes to the 101 nucleotides of the VTE sequence.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01310570
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  • 7
    Electronic Resource
    Electronic Resource
    Deletion analysis of a translational enhancer upstream from the coat protein open reading frame of potato virus S (1997)
    Springer
    Archives of virology 142 (1997), S. 167-175 
    Details
    ISSN: 1432-8798
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary. We have previously demonstrated that the 101 nucleotides upstream from the ATG of the potato virus S (PVS) coat protein gene has the ability to act as a translational enhancer in vitro and in vivo when fused to a range of marker genes. These 101 nucleotides, which have been designated VTE (viral translational enhancer), contain a block of nucleotide homology that is conserved between members of the carlavirus group, centered around a core sequence of CCTTTAGGTT. When deletions were generated that lacked 47 of the 5′ nucleotides, but still retained the conserved block, and analysed in vitro and in vivo, it was observed that these leaders had lost their ability to act as translational enhancers. These results suggest that the sequences 5′ to the conserved block may be acting as a translational enhancer or may be important in placing the conserved block in optimal context. This is confirmed to some extent by hybrid arrest translation results in which the effects on translation of oligonucleotides, complementary to regions within the VTE leader, were investigated. It was observed that oligonucleotides complementary to the nucleotides 5′ to the conserved block had a dramatic effect on the translational competence of transcripts derived from VTE-luciferase constructs, decreasing levels by 53%, whereas oligonucleotides complementary to sequences 3′ to the conserved block reduced levels of translation by only 15.6%.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s007050050067
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  • 8
    Electronic Resource
    Electronic Resource
    Analysis of a translational enhancer present within the 5′-terminal sequence of the genomic RNA of potato virus S (1999)
    Springer
    Archives of virology 144 (1999), S. 1451-1461 
    Details
    ISSN: 1432-8798
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary.  When present as a transcript leader the 5′ untranslated sequence from the potato virus S genomic RNA molecule enhances translation of a downstream open reading frame both in vitro and in vivo. Translational enhancement was 30-fold in rabbit reticulocyte lysate and 15 fold in wheat germ above translation from a transcript with a synthetic leader. Transient expression experiments using tobacco protoplasts and particle bombardment of leaf tissue resulted in enhancement of fourteen and five-fold, respectively, above translation with a synthetic leader. In stably transformed plants the PVS 5′UTR enhanced translation yield ca. 5-fold compared with a synthetic 5′UTR.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s007050050601
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  • 9
    Electronic Resource
    Electronic Resource
    A CaMV 35S promoter driven cDNA clone of tobacco mosaic virus can infect host plant tissue despite being uninfectious when manually inoculated onto leaves (1997)
    Springer
    Archives of virology 142 (1997), S. 183-191 
    Details
    ISSN: 1432-8798
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary. Leading from the success of inoculating plants with viral RNA transcribed in vitro from full length cDNA clones, attempts have been made to build cDNA clones which are directly infectious by inoculation. However, we and others have found that viral cDNA clones driven by the CaMV 35S promoter were able to infect some host plants yet not others, when manually inoculated onto leaves. Alternative methods including microprojectile bombardment have been used to deliver an infectious TMV construct into plant cells resulting in the infection of all TMV host plants tested. Lack of infection via manual inoculation may be due to unsuccessful delivery of a viable construct into the plant cell nucleus.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s007050050069
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  • 10
    Electronic Resource
    Electronic Resource
    A Brassica napus mRNA expressed specifically in developing microspores (1991)
    Springer
    Plant molecular biology 17 (1991), S. 295-299 
    Details
    ISSN: 1573-5028
    Keywords: Brassica napus ; microspore-specific ; microsporogenesis ; DNA sequence ; peptide motif
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The I3 cDNA isolated from a library made from 2–4 mm (immature) anthers of Brassica napus shows microspore-specific expression. Homologous transcripts are detected in buds and anthers of male-fertile plants, but not in green tissues, roots, or in cytoplasmic male-sterile buds. High expression of the transcript is limited to microspores entering and undergoing mitosis. The predicted peptide sequence of the cDNA shows an unusual repeated alanine/proline motif at the C-terminus, which may be of importance in the native protein structure.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00039509
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