molecular cloning, sequencing and expression in escherichia coli of the odontoglossum ringspot virus coat protein gene.the sequence of the 3'-terminal 1865 nucleotides of the genome of the tobamovirus odontoglossum ringspot virus (orsv) was determined. this sequence contained two open reading frames (orfs), 912 and 477 nucleotides long. the 912 nucleotide orf has been identified as the cell-to-cell transport protein gene. the 477 nucleotide orf was expressed in escherichia coli, and the product was detected by antibodies specific for the coat protein of orsv. the amino acid sequence of protein encoded by this or ...19911895062
a dna clone encoding the full-length infectious genome of odontoglossum ringspot tobamovirus and mutagenesis of its coat protein gene.a full-length dna clone encoding the genome of odontoglossum ringspot tobamovirus (orsv) was synthesized and placed adjacent to a bacteriophage t7 rna polymerase promoter. capped-rna transcripts produced in vitro were highly infectious when mechanically inoculated onto seedlings of nicotiana benthamiana and oncidium gower ramsey. a representative clone, designated pot2, caused a disease phenotype identical to that produced by parental viral rna. elisa, western blot analysis, northern blot hybrid ...19989505974
analysis of bovine respiratory syncytial virus envelope glycoproteins in cell compare the requirements for respiratory syncytial virus (rsv)-mediated cell fusion, the fusion (f), attachment (g) and small hydrophobic (sh) glycoproteins of bovine rsv (brsv), ovine rsv (orsv) and human rsv (hrsv) were expressed individually or coexpressed in either homologous or heterologous combinations in hela cells, using the vaccinia virus-t7 polymerase transient expression system. cell fusion was examined by a reporter gene activation assay. although the expression of the f protein a ...19979266983
systemic expression of a bacterial gene by a tobacco mosaic virus-based vector.tobacco mosaic virus (tmv) produces large quantities of rna and protein on infection of plant cells. this and other features, attributable to its autonomous replication, make tmv an attractive candidate for expression of foreign sequences in plants. however, previous attempts to construct expression vectors based on plant rna viruses, such as tmv, have been unsuccessful in obtaining systemic and stable movement of foreign genes to uninoculated leaves in whole plants. a hybrid viral rna (tb2) was ...19911651497
the complete nucleotide sequence of odontoglossum ringspot virus (cy-1 strain) genomic rna.the complete nucleotide sequence of the genomic rna of odontoglossum ringspot virus cy-1 strain (orsv cy-1) was determined using cloned cdna. this sequence is 6611 nucleotides long containing four open reading frames, which correspond to 126 k, 183 k, 31 k, and 18 k proteins. its genomic organization is similar to other tobamoviruses, tmv-v(vulgare), tmv-l (tomato strain), tobacco mild green mosaic virus (tmgmv) and cucumber green mottle mosaic virus (cgmmv). the 5' non-coding regions of orsv cy ...19958789059
tobamovirus evolution: gene overlaps, recombination, and taxonomic implications.tobamoviruses, mostly isolated from solanaceous plants, may represent ancient virus lineages that have codiverged with their hosts. recently completed nucleotide sequences of six nonsolanaceous tobamoviruses allowed assessment of the codivergence hypothesis and support a third subgroup within tobamoviruses. the genomic sequences of 12 tobamoviruses and the partial sequences of 11 others have been analyzed. comparisons of the predicted protein sequences revealed three clusters of tobamoviruses, c ...19968952077
the use of dig-labelled crna probes for the detection of cymbidium mosaic potexvirus (cymmv) and odontoglossum ringspot tobamovirus (orsv) in orchids.dig-labelled sense and antisense crna probes were synthesized from cdna clones of cymmv and orsv for virus detection in infected plants. a slot-blot hybridization assay was developed using either crude leaf extracts or total rna from infected leaves. the assay could detect 50 and 250 pg of purified cymmv and orsv rna, respectively. as little as 30 mg of nicotiana benthamiana infected leaves was sufficient to provide positive detection. cymmv and orsv were detected at 3125 and 625 times dilution ...19989562413
invasion of minor veins of tobacco leaves inoculated with tobacco mosaic virus mutants defective in phloem-dependent fully understand vascular transport of plant viruses, the viral and host proteins, their structures and functions, and the specific vascular cells in which these factors function must be determined. we report here on the ability of various cdna-derived coat protein (cp) mutants of tobacco mosaic virus (tmv) to invade vascular cells in minor veins of nicotiana tabacum l. cv. xanthi nn. the mutant viruses we studied, tmv cp-o, u1mcp15-17, and snc015, respectively, encode a cp from a different t ...19968855325
genomic position affects the expression of tobacco mosaic virus movement and coat protein genes.alterations in the genomic position of the tobacco mosaic virus (tmv) genes encoding the 30-kda cell-to-cell movement protein or the coat protein greatly affected their expression. higher production of 30-kda protein was correlated with increased proximity of the gene to the viral 3' terminus. a mutant placing the 30-kda open reading frame 207 nucleotides nearer the 3' terminus produced at least 4 times the wild-type tmv 30-kda protein level, while a mutant placing the 30-kda open reading frame ...19938446627
the tobamovirus capsid protein functions as a host-specific determinant of long-distance movement.the tobamoviruses, tobacco mosaic virus (tmv) and odontoglossum ringspot virus (orsv), differ in the range of plant species that each can systemically infect. both viruses systemically infect nicotiana benthamiana similarly, but differ in the ability to systemically infect tobacco (n. tabacum). orsv is confined to the inoculated leaves of n. tabacum, whereas tmv causes a rapid systemic infection. genetic chimeras were created to identify viral genes involved in specific interactions in long-dist ...19938438560
type-specific serologic diagnosis of respiratory syncytial virus infection, based on a synthetic peptide of the attachment protein g.peptides deduced from the central hydrophobic region (residues 158-189) of the g protein of bovine and ovine respiratory syncytial virus (rsv) and of human rsv subtypes a and b were synthesized. these peptides were used to develop elisas to measure specifically antibodies against these types and subtypes of rsv. we have evaluated the bovine rsv-g peptide in both an indirect elisa and in a blocking elisa. specificity and sensitivity, relative to a routine diagnostic elisa that detects antibodies ...19968699029
nucleotide and predicted amino acid sequence analysis of the ovine respiratory syncytial virus non-structural 1c and 1b genes and the small hydrophobic protein gene.respiratory syncytial virus (rsv) infects humans and cattle causing serious respiratory tract disease in both. the genome of the human and bovine rsv (hrsv and brsv) codes for two non-structural and eight structural proteins. rsv has also been isolated from naturally infected sheep, but the genome of the ovine rsv (orsv) has not been characterized and nor has the virus host range been identified. here we report on the cloning and sequencing of the two non-structural 1c and 1b genes and of the sm ...19948113762
nucleotide sequence analysis of a cdna clone encoding the 34k movement protein gene of odontoglossum ringspot virus, orsv-cy, the korean isolate.the partial nucleotide sequence of the 3'-terminal region of the korean isolate of odontoglossum ringspot tobamovirus (orsv-cy) from cool-growing cymbidium was determined. the sequence contained a full length open reading frame (orf) coding for the viral cell-to-cell movement protein (mp). the orf was located upstream of the coat protein gene and 105 nucleotides longer than that of tobacco mosaic virus (tmv). the orf predicts a polypeptide chain of 303 amino acids with a molecular weight of 3357 ...19948000012
similarities between the secondary structure of satellite tobacco mosaic virus and tobamovirus rnas.the secondary structure of satellite tobacco mosaic virus (stmv) rna was predicted using computer simulations of rna folding. the analogies of structural elements in the 3' end untranslated regions (3'-utr) of tobamoviral rnas were analysed. in addition to the trna-like structure and pseudoknot stalk, which are found in all known rnas of tobamoviruses and stmv, another region of stable consecutive pseudoknots was predicted in the 3'-utr of stmv rna. a similar pattern of repeated structural units ...19947931178
nucleotide sequence analysis of the non-structural ns1 (1c) and ns2 (1b) protein genes of bovine respiratory syncytial virus.the nucleotide and deduced amino acid sequences of the non-structural (ns) protein genes of bovine respiratory syncytial virus (brsv) strain a51908 were determined. the ns1 and ns2 genes were 524 and 489 nucleotides long with single open reading frames, encoding polypeptides of 136 and 124 amino acids, respectively. comparison of the ns1 gene of brsv with the corresponding sequences of ovine respiratory syncytial virus (orsv) and human respiratory syncytial virus (hrsv) subgroups a and b reveale ...19957844532
an appraisal of the banded and paracrystalline cytoplasmic inclusions induced in cymbidium mosaic potexvirus- and odontoglossum ringspot tobamovirus infected orchid cells using confocal laser scanning microscropy.the cymbidium mosaic potexvirus (cymv) banded inclusions and the odontoglossum ringspot tobamovirus (orsv) paracrystalline inclusions were studied in flowers and leaves of nine orchid cultivars using the confocal laser scanning microscope. the inclusions varied in length and width and were mostly located adjacent to the cell walls. some orsv inclusions fully extended across entire infected cells. we propose that the cymv banded inclusion was formed from virus aggregates which aligned as periodic ...19968634017
rapid detection and identification of odontoglossum ringspot virus by polymerase chain reaction amplification.the odontoglossum ringspot tobamovirus (orsv) movement and coat proteins genes were selected for the design of oligonucleotide primers for amplification of a 1,085 bp fragment. a combined assay of reverse transcription and the polymerase chain reaction (rt-pcr) was performed with 20-mer orsv-specific primers and crude nucleic acid extracts from virus-infected orchids for rapid detection of the virus. the lowest concentration of template viral rna required for detection was 10 fg. the rt-pcr is a ...19958522141
the complete nucleotide sequence and genome organization of odontoglossum ringspot tobamovirus rna.the complete nucleotide sequence of the genomic rna of odontoglossum ringspot tobamovirus (orsv) was determined. the rna genome of orsv is 6618 nucleotides long and contains five open reading frames (orfs 1 to 5) coding for proteins of m(r) 126 k, 181 k, 34 k, 18 k and 52 k, respectively. this is the longest rna of the known viruses of the tobamovirus genus. the sequences of the orsv rna encoded proteins exhibit high homology to the proteins of the members of the tobamovirus genus. the genomic o ...19957487489
[studies on the odontoglossum ringspot virus, a relative of the tobacco mosaic virus. i. physical, chemical, serological and symptomatological findings]. 19654957879
odontoglossum ringspot virus coat protein: sequence and antigenic comparisons with other tobamoviruses.comparative immunochemical analysis of different tobamoviruses indicated that the previously reported coat protein sequence of odontoglossum ringspot virus was likely erroneous. this sequence has been determined again by direct sequencing of the genomic rna and was found to differ from the previously proposed sequence in 31 of the 157 amino acid residues. the extent of antigenic cross-reactivity between orsv protein and other tobamovirus proteins was measured by elisa and found to correlate sati ...19902260922
nucleotide sequence of cell-to-cell transport protein gene of odontoglossum ringspot virus. 19902259639
nucleotide sequence analysis of the ovine respiratory syncytial virus g glycoprotein gene.respiratory syncytial virus (rsv) infects humans and animals including ruminants. among the 10 genes coded for in the viral genome, the putative attachment glycoprotein g gene has been the most variable among strains. human rsv have been divided to two subgroups based on immunological and base sequence data on the attachment glycoprotein g and its gene, respectively. it has been suggested that similar antigenic diversity also exists among bovine rsv (brsv) isolates. in this study, we report on t ...19938372453
synergism in replication of cymbidium mosaic potexvirus (cymmv) and odontoglossum ringspot tobamovirus (orsv) rna in orchid in vitro orchid protoplast isolation method to study replication kinetics of cymmv and orsv was developed. this method allows the isolation of viable and raphid-free petal protoplasts from an orchid hybrid, dendrobium sonia (dendrobium caesar x dendrobium tomie drake). the optimum field strength for both viral rna to achieve good efficiency of electroporation was 750 v/cm and the optimum viral rna concentration required was 1 microgram and 4 micrograms per 2 x 10(6) protoplasts for cymmv and ...19989722873
mutational analysis of the movement protein of odontoglossum ringspot virus to identify a host-range determinant.tobacco mosaic virus (tmv) which contains the movement protein (mp) of odontoglossum ringspot tobamovirus (orsv) in place of the tmv mp systemically infects orchids but causes local infection in tobacco unless the carboxy-terminal 48 amino acids of the mp are deleted (c. a. holt, c. a. fenczik, s. j. casper, and r. n. beachy; virology, in press, 1995). frameshift mutations were created within the 3' ends of the mp gene that led to truncations of the orsv mp by 11, 19, 28, 37, and 48 amino acids; ...19957579612
the gprime package: computer programs for identifying the best regions of aligned genes to target in nucleic acid hybridisation-based diagnostic tests, and their use with plant viruses.the gprime (group primer design) programs examine aligned sets of gene sequences to discover homologous regions to be targeted in diagnostic tests. the core program moves a 'window' over the aligned sequences and calculates, at each window position, a 'redundancy value', namely the number of sequences that would represent all permutations of the variable sequence positions within that window. regions with minimal redundancy values may then be targeted in diagnostic tests based on oligonucleotide ...19989763130
heterologous sequences greatly affect foreign gene expression in tobacco mosaic virus-based vectors.a series of tobacco mosaic virus (tmv)-based hybrid vectors for transient gene expression were constructed with similar designs but differing in the source of heterologous tobamovirus sequence: odontoglossum ringspot virus, tobacco mild green mosaic virus variants u2 and u5, tomato mosaic virus, and sunn-hemp mosaic virus. these vectors contained a heterologous coat protein subgenomic mrna promoter and coat protein open reading frame (orf) and either tmv or heterologous 3' nontranslated region. ...199910069957
expression and characterisation of the ovine respiratory syncytial virus (orsv) g protein for use as a diagnostic reagent.respiratory syncytial virus (rsv) causes severe lower respiratory tract infection in children and calves. antibodies to ovine rsv (orsv) are common in sheep, but the clinical disease is not well defined. there is no report of orsv infection in australian sheep although respiratory distress syndrome has been described. this discrepancy may be due to the lack of a suitable diagnostic test. in this report, we have characterised the orsv g protein in an attempt to study its relatedness to human and ...199910543382
cloning of the 3'-terminal region encoding movement and coat proteins of a korean isolate of odontoglossum ringspot virus.the 3'-terminal 1855 nucleotides (nts) of a korean isolate of odontoglossum ringspot tobamovirus (orsv-cy) were cloned and sequenced. the sequence contained two open reading frames, which encode the cell-to-cell movement protein (mp) and coat protein genes, and are 912 nts and 477 nts long, respectively. the mp gene contained a conserved sequence motif of tobamoviruses and putative assembly origin of the viral rna locating between 1117 nts and 1292 nts from the 3'-end. the 3' untranslated region ...19957733821
rapid simultaneous detection of two orchid viruses using lc- and/or maldi-mass spectrometry.liquid chromatography/mass spectrometry (lc/ms) and matrix-assisted laser desorption-ionization (maldi) mass spectrometry are capable of providing molecular mass information on biological samples with high speed, accuracy and sensitivity. with mass spectrometry, identifying a virus based on the molecular weight of its coat protein is relatively simple and accurate. the technique can be applied to all viruses with known coat protein molecular weights. using the lc/ms and/or maldi, this paper desc ...200010716342
transgenic tobacco plants expressing a coat protein gene of tobacco mosaic virus are resistant to some other tobamoviruses.transgenic tobacco plants expressing the coat protein (cp) gene of tobacco mosaic virus were tested for resistance against infection by five other tobamoviruses sharing 45-82% homology in cp amino acid sequence with the cp of tobacco mosaic virus. the transgenic plants (cp+) showed significant delays in systemic disease development after inoculation with tomato mosaic virus or tobacco mild green mosaic virus compared to the control (cp-) plants, but showed no resistance against infection by ribg ...19902131095
simultaneous td/rt-pcr detection of cymbidium mosaic potexvirus and odontoglossum ringspot tobamovirus with a single pair of primers.cymbidium mosaic potexvirus (cymmv) and odontoglossum ringspot tobamovirus (orsv) are the two most prevalent and economically important orchid viruses which have attained a world-wide distribution. we have designed one pair of common primers, which were able to detect, simultaneously, both viruses by a single polymerase chain reaction (pcr). sequence alignment and primer analyses were employed in primer design. the effect of touch-down pcr, primer annealing temperature, number of thermal cycles, ...19989694327
rescue of a bovine respiratory syncytial virus genomic rna analog by bovine, human and ovine respiratory syncytial viruses confirms the "functional integrity" and "cross-recognition" of brsv cis-acting elements by hrsv and orsv.the nucleotide sequences of the 3' leader and 5' trailer regions were determined for genomic rna of bovine respiratory syncytial virus (brsv) strain a-51908. the leader and trailer sequences are '45' and '161' nucleotides in length, respectively. the functionality of brsv leader and trailer sequences and their recognition by hrsv and ovine respiratory syncytial virus (orsv) proteins were examined with a in vitro transcribed brsv genomic rna analog carrying the bacterial chloramphenicol acetyl tr ...199910550670
the complete sequence of a singapore isolate of odontoglossum ringspot virus and comparison with other tobamoviruses.the complete sequence of a singapore isolate of odontoglossum ringspot virus (orsv-s1) comprises 6609 nucleotides (nt) and four open reading frames (orfs 1 to 4). the 126/183-kda rna-dependent rna polymerase (rdrp), 33-kda movement protein (mp) and 18-kda coat protein (cp) cistrons are located at nt 63-3401/4901, 4807-5718, and 5721-6197 on the genome, respectively. the 5' utr contains three copies of an 8-base direct repeat and (caa)n motifs. characteristic trna-like structure and three consecu ...19968666266
detection of two orchid viruses using quartz crystal microbalance (qcm) immunosensors.quartz crystal microbalance (qcm) immunosensors are based on the principle that adsorption of substances on the surface of a quartz crystal changes its resonance oscillation frequency. a qcm immunosensor was developed for the detection of both cymbidium mosaic potexvirus (cymmv) and odontoglossum ringspot tobamovirus (orsv) by pre-coating the qcms with virus-specific antibodies. upon binding of virions in either purified form or crude sap of infected orchids with the immobilised virus antibodies ...200211684305
mutation of phe50 to ser50 in the 126/183-kda proteins of odontoglossum ringspot virus abolishes virus replication but can be complemented and restored by exact reversion.sequence comparison of a non-biologically active full-length cdna clone of odontoglossum ringspot virus (orsv) pot1 with a biologically active orsv cdna clone pot2 revealed a single nucleotide change of t-->c at position 211. this resulted in the change of phe50 in ot2 to ser50 in ot1. it was not the nucleotide but the amino acid change of phe50 that was responsible for the inability of ot1 to replicate. time-course experiments showed that no minus-strand rna synthesis was detected in mutants wi ...200415269387
simultaneous quantitation of two orchid viruses by the taqman real-time rt-pcr.simultaneous quantitation of two orchid viruses, cymbidium mosaic potexvirus (cymmv) and odontoglossum ringspot tobamovirus (orsv), were carried out using the taqman((r)) real-time rt-pcr, a novel detection technique that combines rt-pcr with the power of fluorescent detection. four taqman((r)) probes were synthesized, targeting at the rna-dependent rna polymerase (rdrp) and coat protein (cp) genes of both viruses. the reporter dye fam (6-carboxyfluorescein) was used to label the 5' terminus of ...200010856762
reciprocal function of movement proteins and complementation of long-distance movement of cymbidium mosaic virus rna by odontoglossum ringspot virus coat protein.complementation of movement and coat proteins of the orchid-infecting potexvirus cymbidium mosaic virus (cymmv) and tobamovirus odontoglossum ringspot virus (orsv) was investigated. nicotiana benthamiana, which is susceptible to both cymmv and orsv, was used as a model system. four transgenic lines, each harbouring one of the movement protein (mp) or coat protein (cp) genes of cymmv or orsv, were constructed. the mp of cymmv consists of three overlapping open reading frames, together called the ...200515831968
expression and purification of a neuropeptide nocistatin using two related plant viral vectors.both odontoglossum ringspot virus (orsv) and tobacco mosaic virus (tmv) were investigated as expression viral vectors for the expression of a neuropeptide nocistatin. chimeras of orsv and tmv were constructed by fusion of 17 amino acids of mouse nocistatin (mnst) to the c-terminal of the coat protein (cp) gene via a factor xa cleavage linker to yield orsv-mnst and tmv-mnst. expression of the mnst peptide was demonstrated by immuno-transmission electron microscopy, western blot, mass spectrometry ...200212036585
significance of the 3'-terminal region in minus-strand rna synthesis of hibiscus chlorotic ringspot virus.rna-dependent rna polymerase (rdrp) was solubilized from crude extracts of hibiscus cannabinus infected by hibiscus chlorotic ringspot virus (hcrsv), a member of the carmoviridae. after treatment of the extracts with micrococcal nuclease to remove the endogenous templates, the full-length genomic rna and the two subgenomic rnas were efficiently synthesized by the partially purified rdrp complex in vitro. when the full-length rnas of potato virus x, tobacco mosaic virus, odontoglossum ringspot vi ...200415166462
detection of two orchid viruses using quartz crystal microbalance-based dna biosensors.abstract we have developed a piezoelectric dna-sensor based on dna-rna hybridization for the detection of two orchid viruses, cymbidium mosaic virus (cymmv) and odontoglossum ringspot virus (orsv). specific oligonucleotide probes modified with a mercaptohexyl group at the 5'-phosphate end were directly immobilized onto 10-mhz at-cut quartz crystal microbalance (qcm). qcms coated with such oligonucleotide probes were exposed to test solutions containing viral rna for hybridization. various experi ...200218944263
molecular beacons: a new approach to plant virus detection.abstract molecular beacons are single-stranded nucleic acid molecules with a stem-loop conformation. the stem portion consists of complementary sequences at the 5' and 3' terminals of the molecule, while the loop portion consists of probe sequences that are complementary to the target sequences of choice. a fluorescent moiety is attached to one end, while a quenching moiety is attached to the opposite end. reverse transcription-polymerase chain reactions are carried out with primers that amplify ...200018944619
detection of cymbidium mosaic potexvirus and odontoglossum ringspot tobamovirus using immuno-capillary zone electrophoresis.abstract immuno-capillary zone electrophoresis (i-cze) is a technique that combines the specificity afforded by serological assays with the sensitivity, rapidity, and automation in detection provided by capillary zone electrophoresis. cymbidium mosaic potexvirus (cymmv) and odontoglossum ringspot tobamovirus (orsv) were detected in their purified forms as well as in the crude saps of infected nicotiana benthamiana leaves and oncidium orchid flowers. the two orchid virus-antibody complexes were r ...199918944725
[molecular identification of cymbidium mosaic potexvirus and odontoglossum ringspot tobamovirus complex infected phalaenopsis and its pathological ultrastructural alteration].filamentous and rod-shaped virions, and aggregated crystals were observed in infected leaves of by negative staining and ultramicrotomy. histologic study synchronously showed typical crystal forms of the two virions: the crystals from filamentous particles congregated in strips, arrayed in multilayer and piled in certain angles or helix between layers; while the crystals from rod-shaped particles arrayed in parallel, angle-layer or helix. the two kinds of crystals both presented in parenchyma ce ...200717580669
cloning and sequencing of coat protein gene of an indian odontoglossum ringspot virus isolate. 200415745051
use of superparamagnetic beads for the isolation of a peptide with specificity to cymbidium mosaic virus.a modified method for the rapid isolation of specific ligands to whole virus particles is described. biopanning against cymbidium mosaic virus was carried out with a commercial 12-mer random peptide display library. a solution phase panning method was devised using streptavidin-coated superparamagnetic beads. the solution based panning method was more efficient than conventional immobilized target panning when using whole viral particles of cymbidium mosaic virus as a target. enzyme-linked immun ...200616781785
virus resistance in orchids.orchid plants, phalaenopsis and dendrobium in particular, are commercially valuable ornamental plants sold worldwide. unfortunately, orchid plants are highly susceptible to viral infection by cymbidium mosaic virus (cymmv) and odotoglossum ringspot virus (orsv), posing a major threat and serious economic loss to the orchid industry worldwide. a major challenge is to generate an effective method to overcome plant viral infection. with the development of optimized orchid transformation biotechnolo ...201425438783
detection of viruses directly from the fresh leaves of a phalaenopsis orchid using a microfluidic system.early detection of pathogens is crucial for the effective surveillance of diseases. many efforts have been made to explore methods which can detect these pathogens within a short period of time without requiring a tedious protocol. however, these developed methods have disadvantages such as they are relatively time-consuming or require specialized laboratory facilities. in this work, we have developed an integrated microfluidic system for rapid and automatic detection of viruses by direct analys ...201323751373
using ac-field-induced electro-osmosis to accelerate biomolecular binding in fiber-optic sensing chips with microstructures.this article reports the use of ac-field-induced charges at the corners of microstructures on fiber-optic sensing chips to generate electro-osmotic vortex flows in flow cell channels that can accelerate solute binding on the fiber. the sensing chip made of a cyclic olefin copolymer coc substrate contained a flow cell channel of dimensions 15 mm x 1 mm x 1 mm. a partially unclad optical fiber was placed within the channel. relief-like strip structures of 25-mum thickness fabricated on the channel ...201020055421
multiplex rt-pcr detection of three common viruses infecting orchids.a multiplex reverse transcription polymerase chain reaction (rt-pcr) assay was developed for simultaneous detection of three orchid viruses: cymbidium mosaic virus (cymmv), odontoglossum ringspot virus (orsv), and orchid fleck virus (ofv). primers were used to amplify nucleocapsid protein gene fragments of 845 bp (orsv), 505 bp (cymmv) and 160 bp (ofv). a 60-bp amplicon of plant glyceraldehyde-3-phophate dehydrogenase mrna was included as an internal control against false negatives. the assay wa ...201424980395
odontoglossum ringspot virus host range restriction in nicotiana sylvestris maps to the replicase gene.summary the experimental host range of odontoglossum ringspot virus (orsv), a member of the tobamoviruses, includes several species of nicotiana, but not n. sylvestris. however, orsv was able to replicate in protoplasts from n. sylvestris leaves. by using the green fluorescent protein (gfp) as a marker inserted into orsv, it was found that a small number of single epidermal cells became infected in mechanically inoculated leaves, but the virus did not move cell to cell. the orsv movement protein ...200520565669
simultaneous detection of cymbidium mosaic virus and odontoglossum ringspot virus in orchids using multiplex rt-pcr.a system for simultaneous detection of two orchid-infecting viruses was developed and applied to several orchid species. the detection system involved multiplex reverse transcription-polymerase chain reaction (rt-pcr) and could simultaneously identify cymbidium mosaic virus (cymmv) and odontoglossum ringspot virus (orsv) from the orchid species studied. multiplex rt-pcr was conducted using two virus-specific primer pairs and an internal control pair of primers to amplify the cymmv and orsv coat ...201526542829
direct detection of orchid viruses using nanorod-based fiber optic particle plasmon resonance immunosensor.a fiber optic particle plasmon resonance (foppr) immunosensor is developed for label-free detection of orchid viruses that use gold nanorods (aunrs) as the sensing material. the aunrs are employed to create a near-infrared sensing window to solve the color interference problem of sample matrix for direct sensing of target analyte. this work cannot be achieved using gold nanospheres (aunss) because the signal of sample color absorption largely overlaps the signal of molecular recognition events i ...201424001513
characterization and application of a common epitope recognized by a broad-spectrum c4 monoclonal antibody against capsid proteins of plant potyviruses.a broad-spectrum monoclonal antibody (c4 mab) against the capsid proteins (cps) of plant potyviruses has been generated in previous studies. to clarify which epitope is recognized by this mab, epitope mapping was performed via phage display library screening and amino acid substitution analysis. subsequently, a 12-residue epitope in the core region of potyvirus cps was identified and termed the c4 epitope (wxmmdgxxqxxy/f). this epitope contains tryptophan and tyrosine residues that are crucial f ...201626541335
genetic variability in the coat protein genes of two orchid viruses: cymbidium mosaic virus and odontoglossum ringspot virus.the variability in coat protein gene sequences of cymbidium mosaic virus (cymmv) and odontoglossum ringspot virus (orsv) that naturally infect orchids worldwide was investigated. samples were collected from korea, singapore and taiwan. the sequence data were compared with available published coat protein gene sequences of cymmv and orsv, including those from japan and thailand. among cymmv isolates, the homology was 89.1%-99.7% and 93.2%-100% at the nucleotide and amino acid levels, respectively ...200212376755
application of an integrated omics approach for identifying host proteins that interact with odontoglossum ringspot virus capsid protein.the glutamic acid at position 100 (e(100)) in the capsid protein (cp) of odontoglossum ringspot virus (orsv) plays an important role in long-distance viral movement in nicotiana benthamiana. the orsv(e100a) mutant, which has a glutamic acid to alanine substitution, shows a loss of systemic infectivity in n. benthamiana. transmission electron microscopy and size-exclusion chromatography assays showed that e(100) is essential for cp-cp interaction and viral particle assembly. to identify the orsv ...201525625820
monoclonal antibody-based triple antibody sandwich-enzyme-linked immunosorbent assay and immunocapture reverse transcription-polymerase chain reaction for odontoglossum ringspot virus detection.odontoglossum ringspot virus (orsv) infects numerous commercially important orchids and causes significant losses worldwide. the coat protein (cp) gene of orsv was cloned and expressed in escherichia coli by using the pet-32a expression vector, and the expression of recombinant protein was confirmed by western blotting using anti-orsv antibodies. the recombinant protein was purified using ni-nta agarose, and the purified protein was used as an immunogen to produce monoclonal antibodies (mabs) an ...201020933014
high sequence conservation among odotoglossum ringspot virus isolates from orchids.the variability in the nucleotide (nt) and amino acid (aa) sequences of the coat protein (cp) of odontoglossum ringspot virus (orsv), which naturally infects orchids worldwide, was investigated. the cp genes of 48 orsv isolates originating from different locations in korea were amplified using rt-pcr and sequenced. the encoded cp consists of 158 aa. the cp sequences of the korean isolates were compared at the nt and aa levels with those of the previously published orsv isolates originating from ...201021127958
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