development of a chromatographic strip assay for detection of porcine antibodies to 3abc non-structural protein of foot-and-mouth disease virus serotype o.a chromatographic strip assay was developed for rapid detection of serum antibodies to non-structural protein of foot-and-mouth disease virus. the assay was based on escherichia coli-expressed 3abc non-structural protein and an immunochromatographic technique, which shortened the detection time to about one hour. the sensitivity of the assay was determined to be 96.8% for infected pigs; its specificity was 100% for naïve pigs and 98.8% for vaccinated pigs. in the experimentally infected pigs, an ...200919578276
hydrolytic properties and substrate specificity of the foot-and-mouth disease leader protease.foot-and-mouth disease virus, a global animal pathogen, possesses a single-stranded rna genome that, on release into the infected cell, is immediately translated into a single polyprotein. this polyprotein product is cleaved during synthesis by proteinases contained within it into the mature viral proteins. the first cleavage is performed by the leader protease (lb(pro)) between its own c-terminus and the n-terminus of vp4. lb(pro) also specifically cleaves the two homologues of cellular eukaryo ...200919580333
a long-range rna-rna interaction between the 5' and 3' ends of the hcv genome.the rna genome of the hepatitis c virus (hcv) contains multiple conserved structural cis domains that direct protein synthesis, replication, and infectivity. the untranslatable regions (utrs) play essential roles in the hcv cycle. uncapped viral rnas are translated via an internal ribosome entry site (ires) located at the 5' utr, which acts as a scaffold for recruiting multiple protein factors. replication of the viral genome is initiated at the 3' utr. bioinformatics methods have identified oth ...200919605533
expression of exogenous ifn-alpha by bypassing the translation block protects cells against fmdv infection.foot-and-mouth disease virus (fmdv) is the most contagious pathogen of cloven-hoofed animals. previous studies have demonstrated that type i interferons [alpha/beta interferons (ifn-alpha/betas)] can suppress fmdv replication and spread. conversely, fmdv can also inhibit ifn-alpha expression in infected cells by blocking cap-dependent translation. to overcome the blockade on ifn-alpha mrna translation during fmdv infection, we generated an ires-ifn construct that carries fmdv's internal ribosome ...200919607862
multiple origins of foot-and-mouth disease virus serotype asia 1 outbreaks, 2003-2007.we investigated the molecular epidemiology of foot-and-mouth disease virus (fmdv) serotype asia 1, which caused outbreaks of disease in asia during 2003-2007. since 2004, the region affected by outbreaks of this serotype has increased from disease-endemic countries in southern asia (afghanistan, india, iran, nepal, pakistan) northward to encompass kyrgyzstan, tajikistan, uzbekistan, several regions of the people's republic of china, mongolia, eastern russia, and north korea. phylogenetic analysi ...200919624919
bypass suppression of small-plaque phenotypes by a mutation in poliovirus 2a that enhances apoptosis.the rate of protein secretion in host cells is inhibited during infection with several different picornaviruses, with consequences likely to have significant effects on viral growth, spread, and pathogenesis. this sin(+) (secretion inhibition) phenotype has been documented for poliovirus, foot-and-mouth disease virus, and coxsackievirus b3 and can lead to reduced cell surface expression of major histocompatibility complex class i and tumor necrosis factor receptor as well as reduced extracellula ...200919625405
systematic study of the genetic response of a variable virus to the introduction of deleterious mutations in a functional capsid region.we have targeted the intersubunit interfaces in the capsid of foot-and-mouth disease virus to investigate the genetic response of a variable virus when individual deleterious mutations are systematically introduced along a functionally defined region of its genome. we had previously found that the individual truncation (by mutation to alanine) of 28 of the 42 amino acid side chains per protomer involved in interactions between capsid pentameric subunits severely impaired infectivity. we have now ...200919625409
[construction and identification of recombinant bhv-1 expressing foot and mouth disease virus vp1 gene].in order to construct the recombinant bovine hepervirus-1 (bhv-1) which expressed foot and mouth disease virus (fmdv) vp1 gene, we constructed a bhv-1 ge gene transfer vector by inserting the synthetic vp1 gene of fmdv (o/china/99) under the immediate-early promoter of cytomegalovirus.200919637579
[construction and immunogenicity of recombinant adenovirus co-expressing the gp5 and m protein of porcine reproduction and respriratory syndrome virus in mice].fmdv 2a peptide was introduced as a linker between gp5 and m protein of porcine reproduction and respiratory syndrome virus (prrsv) to allow automatic self-cleavage the polyproteins. this strategy simultaneously displayed the neutralizing action of gp5 protein and cell-mediated immunity of m protein. we put them into the expression cassette of adenovirus vector. the results of rt-pcr, ifa and western blotting showed that gp5 and m protein were not only expressed correctly, but also self-cleavage ...200919637620
[molecular design and immunogenicity of a multiple-epitope foot-and-mouth disease virus antigen, adjuvants, and dna vaccination].we designed and constructed a fuse expression gene oaat and staphylococcal enterotoxin a (sea) on the basis of the oaat designed and constructed which consists of the structural protein vp1 genes from serotypes a and o fmdv, 5 major vp1 immunodominant epitopes from two genotypes of asia1 serotype, and 3 th2 epitopes originating from the non-structural protein, 3abc gene and structural protein vp4 gene. the recombinant plasmids pea was constructed using sea as a genetic adjuvant. expressions of t ...200919637624
[fusion expression of escherichia coli heat-labile enterotoxin b subunit gene and foot-and-mouth disease virus type o vp1 gene and immunogenicity analysis].ltb gene fragment was amplified by pcr from plasmid pmdtlt, and a recombinant plasmid petltbvp1 was constructed by inserting ltb gene fragment into vp1 gene expression plasmid petvp1 constructed previously. the recombinant plasmids were transformed into e. coli bl21(de3) and induced to express by iptg. the recombinant protein existed in the inclusion body and its molecular weight was about 39 kd proved by sds-page analysis. western blotting showed that the fusion protein could be reacted with bo ...200919637632
highly sensitive fetal goat tongue cell line for detection and isolation of foot-and-mouth disease virus.a fetal goat cell line (zz-r 127) supplied by the collection of cell lines in veterinary medicine of the friedrich loeffler institute was examined for susceptibility to infection by foot-and-mouth disease (fmd) virus (fmdv) and by two other viruses causing clinically indistinguishable vesicular conditions, namely, the viruses of swine vesicular disease and vesicular stomatitis. primary bovine thyroid (bty) cells are generally the most sensitive cell culture system for fmdv detection but are prob ...200919656987
a multicistronic dna vaccine induces significant protection against tuberculosis in mice and offers flexibility in the expressed antigen repertoire.concerns about the safety and efficacy of mycobacterium bovis bacillus calmette-guérin (bcg) emphasize the need for alternative tuberculosis (tb) vaccines. dna vaccines are interesting candidates but are limited by the restricted antigen repertoire that they express. traditional polycistronic vectors are large and have imbalanced expression. recent advances in molecular genetics and cellular immunology have paved the way toward the rational design of an efficacious vaccine. we exploited self-cle ...200919656992
[establishment of colloidal gold-immunochromatography assay strip for detection of type asia1 foot-and-mouth disease virus].to establish a sensitive, rapid and simple gold immunochromatography assay (gica) for detecting asia1 type of foot-and-mouth disease virus (fmdv) from the field samples. the purified anti-fmdv type asia1 monoclonal antibody labeled with colloidal gold and the goat anti-guinea pig igg were wrapped onto nitrocellulose membrane as the test line (t line) and the control line (c line), respectively. the strip was then further optimized. a total of 87 field samples were detected. the results indicated ...200919670648
production of fmdv virus-like particles by a sumo fusion protein approach in escherichia coli.virus-like particles (vlps) are formed by the self-assembly of envelope and/or capsid proteins from many viruses. some vlps have been proven successful as vaccines, and others have recently found applications as carriers for foreign antigens or as scaffolds in nanoparticle biotechnology. however, production of vlp was usually impeded due to low water-solubility of recombinant virus capsid proteins. previous studies revealed that virus capsid and envelope proteins were often posttranslationally m ...200919671144
[establishment of rt- lamp for rapid detection of foot-and-mouth disease virus].a rapid detection of foot-and-mouth disease virus (fmdv) was established by using reverse transcription loop-mediated isothermal amplification (rt-lamp) method, meanwhile its specificity and sensitivity were assessed. the results showed that the fmdv rna could be amplified by incubation at 65degrees c for only 1h using six primers designed based on fmdv polyprotein gene and the amplification products could be detected easily by naked-eye. there is no cross reaction with other virus such as svdv, ...200919678569
options for control of foot-and-mouth disease: knowledge, capability and policy.foot-and-mouth disease can be controlled by zoo-sanitary measures and vaccination but this is difficult owing to the existence of multiple serotypes of the causative virus, multiple host species including wildlife and extreme contagiousness. although intolerable to modern high-production livestock systems, the disease is not usually fatal and often not a priority for control in many developing countries, which remain reservoirs for viral dissemination. phylogenetic analysis of the viruses circul ...200919687036
extension of flavivirus protein c differentially affects early rna synthesis and growth in mammalian and arthropod host cells.the translation of flaviviral rna genomes yields a single polyprotein that is processed into the mature proteins by viral and host cell proteases. mature capsid protein c is freed from the polyprotein by the viral ns2b/3 protease, cleaving in the c-terminal region of protein c in front of the signal sequence for prm. protein c has been shown to be involved in viral assembly and rna packaging. to examine further the role of protein c and its production by proteolysis, we replaced the ns2b/3 capsi ...200919692461
screening and identification of b cell epitopes of structural proteins of foot-and-mouth disease virus serotype asia1.the objective of this study was to screen and identify the b cell epitopes of structural proteins of foot-and-mouth disease virus (fmdv) serotype asia1. the complete amino acid sequence of all the four structural proteins (p1 region) was analyzed using the dnastar protean system. seventeen peptides were predicted and selected as potential b cell epitopes. the potential b cell epitope genes were cloned into the pgex-6p-1 plasmid, then expressed and purified. the resulting 17 glutathione s-transfe ...201019699594
foot-and-mouth disease virus assembly: processing of recombinant capsid precursor by exogenous protease induces self-assembly of pentamers in vitro in a myristoylation-dependent manner.the assembly of foot-and-mouth disease virus (fmdv) particles is poorly understood. in addition, there are important differences in the antigenic and receptor binding properties of virus assembly and dissociation intermediates, and these also remain unexplained. we have established an experimental model in which the antigenicity, receptor binding characteristics, and in vitro assembly of capsid precursor can be studied entirely from purified components. recombinant capsid precursor protein (p1 r ...200919710148
identification of rna helicase a as a new host factor in the replication cycle of foot-and-mouth disease virus.foot-and-mouth disease virus (fmdv), as with other rna viruses, recruits various host cell factors to assist in the translation and replication of the virus genome. in this study, we investigated the role of rna helicase a (rha) in the life cycle of fmdv. immunofluorescent microscopy (ifm) showed a change in the subcellular distribution of rha from the nucleus to the cytoplasm in fmdv-infected cells as infection progressed. unlike nuclear rha, the rha detected in the cytoplasm reacted with an an ...200919710149
different infection parameters between dairy cows and calves after an infection with foot-and-mouth disease virus.clinical observations of a foot-and-mouth disease (fmd) virus infection in dairy cows and calves were different. this raised the question whether they would also differ with respect to virus excretion and transmission. data were available from transmission experiments carried out with groups of dairy cows and calves. half of each group was inoculated with fmdv o/ned/2001; the other half contact-exposed to inoculated animals. virus excretion, clinical signs and antibody response were measured and ...201019716326
simple and rapid lateral-flow assay for the detection of foot-and-mouth disease virus.a simple lateral-flow assay (lfa) based on a monoclonal antibody (mab 70-17) was developed for the detection of foot-and-mouth disease virus (fmdv) under nonlaboratory conditions. the lfa was evaluated with epithelial suspensions (n = 704) prepared from current and historical field samples which had been submitted to the pirbright laboratory (united kingdom) and from negative samples (n = 100) collected from naïve animals in korea. four fmdv serotypes (type o, a, asia 1, and c) were detected in ...200919726619
structure and dynamics of the gh loop of the foot-and-mouth disease virus capsid.the gh loop of vp1 of the foot-and-mouth disease virus capsid is important because it is a major antigenic site and an integrin recognition site. the gh loop is disordered in all x-ray structures of the capsid except for serotype o under reduced conditions in which the loop lies on the capsid surface. although the structure of the capsid-integrin complex has not yet been determined, the gh loop is known to protrude from the capsid surface when the capsid is bound with an antigen-binding fragment ...200919734079
[construction of eukaryotic expression vector containing foot-and-mouth disease virus vp1 gene and its expression in dendritic cells].to construct the eukaryotic expression vectors for the fmdv vp1 gene and transfect dendritic cells(dc) for the expression of vp1.200919737465
investigations into the cause of foot-and-mouth disease virus seropositive small ruminants in cyprus during 2007, serological evidence for foot-and-mouth disease (fmd) infection was found as a result of differential diagnostic testing of cypriot sheep suspected to be infected with bluetongue or contagious ecthyma. seropositive sheep and goats were subsequently uncovered on ten geographically clustered flocks, while cattle and pigs in neighbouring herds were all seronegative. these antibodies were specific for serotype-o fmd virus, reacting with both structural and non-structural (ns) fmd viral prot ...200919744234
phenotypic and functional characterization of t-cells and in vitro replication of fmdv serotypes in bovine lymphocytes.peripheral blood mononuclear cells (pbmcs) from unvaccinated and vaccinated bovine calves were employed to study the alteration of different t-cell subpopulations, functional competence and detection of foot-and-mouth disease virus (fmdv) at different hours post in vitro infection (hpi) with fmdv serotype either o or a or asia1. the alteration in t-lymphocyte subpopulations was analyzed by flow cytometry analysis using t-cell subpopulation specific monoclonal antibodies at various hpi. all the t ...200919751690
multiplex primer prediction software for divergent targets.we describe a multiplex primer prediction (mpp) algorithm to build multiplex compatible primer sets to amplify all members of large, diverse and unalignable sets of target sequences. the mpp algorithm is scalable to larger target sets than other available software, and it does not require a multiple sequence alignment. we applied it to questions in viral detection, and demonstrated that there are no universally conserved priming sequences among viruses and that it could require an unfeasibly lar ...200919759213
foot-and-mouth disease virus antigen detection enzyme-linked immunosorbent assay using multiserotype-reactive monoclonal antibodies.monoclonal antibody (mab)-based sandwich direct enzyme-linked immunosorbent assay (msd-elisa) methods that can detect foot-and-mouth disease virus (fmdv) antigens, both multiserotype (msd-elisa/ms) (for o, a, c, and asia 1) and single-serotype (msd-elisa/ss) (for o, a, and asia 1, specifically), were developed. mab 1h5 was used as an antigen-trapping antibody that reacted with all seven serotypes of fmdv. the mabs 71f2, 70c4, 16c6, and 7c2 were used as peroxidase-labeled detecting antibodies for ...200919759230
genetic characterization of foot-and-mouth disease viruses, ethiopia, 1981-2007.foot-and-mouth disease (fmd) is endemic to sub-saharan africa. to further understand its complex epidemiology, which involves multiple virus serotypes and host species, we characterized the viruses recovered from fmd outbreaks in ethiopia during 1981-2007. we detected 5 of the 7 fmdv serotypes (o, a, c, southern african territories [sat] 1, and sat 2). serotype o predominated, followed by serotype a; type c was not recognized after 1983. phylogenetic analysis of virus protein 1 sequences indicat ...200919788808
rna nuclear export is blocked by poliovirus 2a protease and is concomitant with nucleoporin cleavage.cytopathic viruses have developed successful strategies to block or, at least, to attenuate host interference with their replication. here, we have analyzed the effects of poliovirus 2a protease on rna nuclear export. 2a protease interferes with trafficking of mrnas, rrnas and u snrnas from the nucleus to the cytoplasm, without any apparent effect on trna transport. traffic of newly produced mrnas is more strongly affected than traffic of other mrnas over-represented in the cytoplasm, such as mr ...200919789179
interaction energy analysis of peptide can predict the possibilities of mimetics by its retroinverso has been previously reported that the retroinverso analog of s peptide cannot mimic the s peptide, whereas the retroinverso analog of foot-and-mouth disease virus antigen can mimic the foot-and-mouth disease virus antigen. the structures of s peptide, foot-and-mouth disease virus antigen, and their retroinverso analogs are known. here, we have attempted to explain the structural basis of mimetics at the level of atomic interactions by elaborating upon the guptasarma's hypothesis. using intera ...200919811507
development and laboratory validation of a lateral flow device for the detection of serotype sat 2 foot-and-mouth disease viruses in clinical samples.a lateral flow device (lfd) for the detection of foot-and-mouth disease virus (fmdv) of the sat 2 serotype was developed using a monoclonal antibody (mab 2h6). the performance of the lfd was evaluated in the laboratory on suspensions of vesicular epithelia: 305 positive for fmdv type sat 2 from suspected cases of vesicular disease collected from 30 countries and 1002 samples shown to be negative for fmdv type sat 2 collected from 67 countries between 1968 and 2008. the diagnostic sensitivity of ...201019815031
equine rhinitis a virus and its low ph empty particle: clues towards an aphthovirus entry mechanism?equine rhinitis a virus (erav) is closely related to foot-and-mouth disease virus (fmdv), belonging to the genus aphthovirus of the picornaviridae. how picornaviruses introduce their rna genome into the cytoplasm of the host cell to initiate replication is unclear since they have no lipid envelope to facilitate fusion with cellular membranes. it has been thought that the dissociation of the fmdv particle into pentameric subunits at acidic ph is the mechanism for genome release during cell entry, ...200919816570
development and laboratory evaluation of a lateral flow device for the detection of swine vesicular disease virus in clinical samples.a lateral flow device (lfd) for the detection of swine vesicular disease (svd) virus (svdv) and differential diagnosis from foot-and-mouth disease (fmd) was developed using a monoclonal antibody (mab c70). the performance of the lfd was evaluated in the laboratory on suspensions of vesicular epithelia and cell culture passage derived supernatants of svdv and porcine teschovirus (enterovirus; pev). the collection of test samples included 157 which were positive for svdv (84 vesicular epithelial s ...201019819260
detection of asymptomatic antigenemia in pigs infected by porcine reproductive and respiratory syndrome virus (prrsv) by a novel capture immunoassay with monoclonal antibodies against the nucleocapsid protein of prrsv.routine surveillance for porcine reproductive and respiratory syndrome virus (prrsv) infections is crucial for the epidemiological control of this disease. antibody tests are widely used but cannot differentiate between vaccination and reinfection. we developed a prrsv antigen capture enzyme-linked immunosorbent assay (elisa) using well-characterized monoclonal antibodies (mabs) raised against the nucleocapsid (n) protein of north american and european prrsv. this antigen assay detected purified ...200919828768
natural killer cell dysfunction during acute infection with foot-and-mouth disease virus.natural killer (nk) cells provide one of the initial barriers of cellular host defense against pathogens, in particular intracellular pathogens. the role of these cells in foot-and-mouth disease virus (fmdv) infection is unknown. previously, we characterized the phenotype and function of nk cells from swine (f. n. toka et al., j. interferon cytokine res. 29:179-192, 2009). in the present study, we report the analysis of nk cells isolated from animals infected with fmdv and tested ex vivo and sho ...200919828769
immune response and viral persistence in indian buffaloes (bubalus bubalis) infected with foot-and-mouth disease virus serotype asia 1.despite their potential role in the spread of foot-and-mouth disease (fmd), the immune response and viral persistence in fmd virus (fmdv)-infected indian buffaloes (bubalus bubalis) have been unexplored. we found similar kinetics of neutralizing antibody responses in the sera and secretory fluids of buffaloes following experimental fmdv asia 1 infection, but the lymphocyte-proliferative response in infected buffaloes was of low magnitude. despite inducing a significant systemic and secretory imm ...200919828770
bola-dr peptide binding pockets are fundamental for foot-and-mouth disease virus vaccine design in cattle.this large study investigates the associations between bovine major histocompatibility complex drb3 alleles and their binding pockets with the immune response to a 40-mer peptide derived from foot-and-mouth disease virus (fmdv) vp1. a crossbred (charolais and holstein) cattle population (n=197) was immunised with the fmdv peptide and specific igg1 and igg2 responses were measured. eighteen different drb3 alleles were detected in this population, with several exhibiting highly significant associa ...200919833250
[construction of an infectious cdna clone derived from foot-and-mouth disease virus o/qyys/s/06].after sequencing, we amplified and cloned foot-and-mouth disease virus (fmdv) o/qyys/s/06 whole genome by three fragments. these three fragments were cloned into vector p43 one by one to construct recombinant plasmid p43c, which carried the full-length cdna of fmdv o/qyys/s/06. then, plasmid p43c and plasmid t7 expressing t7 rna polymerase were co-transfected into bhk-21 cells. after 48 h, we harvested the culture broth from transfected bhk-21 cells and inoculated into 2-3 day-old sucking mice. ...200919835137
foot and mouth disease virus vaccines.foot and mouth disease (fmd) is a highly infectious and economically devastating disease of livestock. although vaccines, available since the early 1900s, have been instrumental in eradicating fmd from parts of the world, the disease still affects millions of animals around the globe and remains the main sanitary barrier to the commerce of animals and animal products. currently available inactivated antigen vaccines applied intramuscularly to individual animals, confer serotype and subtype speci ...200919837296
differential ifn-alpha/beta production suppressing capacities of the leader proteins of mengovirus and foot-and-mouth disease virus.picornaviruses encompass a large family of rna viruses. some picornaviruses possess a leader (l) protein at the n-terminus of their polyprotein. the l proteins of encephalomyocarditis virus, a cardiovirus, and foot-and-mouth disease virus (fmdv), an aphthovirus, are both dispensable for replication and their major function seems to be the suppression of antiviral host cell responses. previously, we showed that the l protein of mengovirus, a strain of encephalomyocarditis virus, inhibits antivira ...201019863558
studies on the physical and chemical properties of the virus of foot-and-mouth disease : i. description of the strain of virus used. titration and centrifugation experiments.a strain of foot-and-mouth disease virus was recovered from a cow at the height of the disease, and was propagated through at least 261 passages in the guinea pig. considerably over 2000 animals proved susceptible to the virus, and the virus could be transferred at will back to cattle and hogs, and then again returned to guinea pigs. no natural immunity was discovered in guinea pigs. secondary lesions were easily and regularly induced, thus making this strain particularly favorable to experiment ...192719869281
studies on the physical and chemical properties of the virus of foot-and-mouth disease : ii. cataphoresis and filtration.cataphoresis experiments show that the virus of foot-and-mouth disease carries, under ordinary conditions, an electropositive charge. its isoelectric range is at the high point of ph = about 8. although cultivable bacteria are, as a rule, electronegative, yet certain protozoa, such as trypanosomes and spirochetes are also electropositive. in respect to charge, then, the virus is different from ordinary bacteria, but there is nothing in this finding to indicate an inanimate character of the incit ...192719869282
statistical evaluation of sieve constants in ultrafiltration.the partial retention of the disperse phase in the ultrafiltration of a monodisperse system through an isoporous filter is interpreted on a statistical basis, and a simple expression for the sieve constant is evaluated in terms of the calibrated membrane porosity and the particle size. curves calculated from this expression are in reasonable agreement with experimental data for the ultrafiltration of serum albumin, hemocyanin (helix), and foot-and-mouth disease virus.193619872986
[rescue and identification foot-and-mouth disease virus asia1/js/china/2005 strain with arg-gly-asp rgd receptor recognition site].to construct an infectious full-length cdna clone of asial/js/china/2005 strain with arg-gly-asp (rgd) receptor recognition site.200919873760
insights into cleavage specificity from the crystal structure of foot-and-mouth disease virus 3c protease complexed with a peptide substrate.picornavirus replication is critically dependent on the correct processing of a polyprotein precursor by 3c protease(s) (3c(pro)) at multiple specific sites with related but non-identical sequences. to investigate the structural basis of its cleavage specificity, we performed the first crystallographic structural analysis of non-covalent complexes of a picornavirus 3c(pro) with peptide substrates. the x-ray crystal structure of the foot-and-mouth disease virus 3c(pro), mutated to replace the cat ...201019883658
differences in the virulence of two strains of foot-and-mouth disease virus serotype a with the same spatiotemporal distribution.during the 2000-2001 epidemic of foot-and-mouth disease virus (fmdv) in argentina, two fmdv serotype a viruses were identified among others. since different pathogenic properties between these virus strains were noticed in cattle, we evaluated several biological properties and features of fmdv a/arg/00 and fmdv a/arg/01 in order to compare these viruses in terms of virulence and pathogenicity. our results indicate that fmdv a/arg/00 grows less efficiently than fmdv a/arg/01, exemplified by small ...201019883705
the 5'-untranslated region of the mouse mammary tumor virus mrna exhibits cap-independent translation this study, we demonstrate the identification of an internal ribosome entry site (ires) within the 5'-untranslated region (5'-utr) of the mouse mammary tumor virus (mmtv). the 5'-utr of the full-length mrna derived from the infectious, complete mmtv genome was cloned into a dual luciferase reporter construct containing an upstream renilla luciferase gene (rluc) and a downstream firefly luciferase gene (fluc). in rabbit reticulocyte lysate, the mmtv 5'-utr was capable of driving translation of ...201019889724
use of a baculovirus-expressed structural protein for the detection of antibodies to foot-and-mouth disease virus type a by a blocking enzyme-linked immunosorbent assay.a blocking enzyme-linked immunosorbent assay (elisa) with a baculovirus-expressed structural protein was developed for the detection of antibodies to foot-and-mouth disease virus type a. it exhibited 99% specificity with a cutoff of 53% inhibition. its sensitivity was comparable to the sensitivities of the virus neutralization test and the liquid-phase blocking elisa, indicating its potential as an alternative assay.201019889938
generation of full-length functional antibody against pres2 of hepatitis b virus in hepatic cells in vitro from bicistrons mediated by gutless adenovirus.monoclonal antibodies (mabs) have been developed as effective therapeutics for a wide variety of diseases. delivery of mabs by gene transfer provides an option for overcoming the difficulties in mab production and manufacturing processes. however, for the polymeric structure of full-length mabs, it is important to design an optimal gene transfer system for mab generation.200919894780
the region between the two polyprotein initiation codons of foot-and-mouth disease virus is critical for virulence in explore the role in viral pathogenesis of the region located between the two functional aug (inter-aug) in foot-and-mouth disease virus (fmdv), we derived viruses containing transposon (tn) inserts from a mutagenized cdna infectious clone of fmdv (pa24-wt). mutant viruses containing an in-frame 57-nt transposon insertion grew at a slower rate and had a smaller plaque size phenotype than the parental virus (a24-wt). a mutant virus containing a 51-nt deletion in inter-aug had a similar phenotyp ...201019896688
effect of fmd vaccine antigen payload on protection, sub-clinical infection and persistence following needle challenge in sheep.the relationship of foot-and-mouth disease (fmd) virus antigen payload and single and double vaccinations in conferring protection against virus challenge in sheep was studied. sheep vaccinated with half the cattle dose (1 ml) containing 15 and 3.75 μg of fmdv antigen with or without booster resisted virulent challenge on 21 days post vaccination or 7 days post booster. fmdv rna could be detected in nasal secretions in 26% of vaccinated sheep (10(3.12) to 10(3.82) viral rna copies) on day 35 pos ...201019896714
optimization of immunohistochemical and fluorescent antibody techniques for localization of foot-and-mouth disease virus in animal tissues.immunohistochemical (ihc) and fluorescent antibody (fa) techniques were optimized for the detection of foot-and-mouth disease virus (fmdv) structural and nonstructural proteins in frozen and paraformaldehyde-fixed, paraffin-embedded (pfpe) tissues of bovine and porcine origin. immunohistochemical localization of fmdv was compared with 7 detection systems, 8 primary antibodies, and 11 epitope retrieval techniques. all serotypes tested (o, a, asia, c [cryosection]; o, a, asia [pfpe]) were localize ...200919901278
prevalence estimates of antibodies towards foot-and-mouth disease virus in small ruminants in uganda.foot-and-mouth disease (fmd) is endemic in uganda with control strategies focusing on vaccination of cattle, while small ruminants are largely ignored. in order for uganda to establish effective control strategies, it is crucial that the epidemiology of the disease is fully understood. this study summarizes results of serological investigations of sheep and goats for antibodies to fmdv from four districts in 2006 following an fmd outbreak in the region and from an attempted comprehensive random ...200919909475
potential benefits of sequential inhibitor-mutagen treatments of rna virus infections.lethal mutagenesis is an antiviral strategy consisting of virus extinction associated with enhanced mutagenesis. the use of non-mutagenic antiviral inhibitors has faced the problem of selection of inhibitor-resistant virus mutants. quasispecies dynamics predicts, and clinical results have confirmed, that combination therapy has an advantage over monotherapy to delay or prevent selection of inhibitor-escape mutants. using ribavirin-mediated mutagenesis of foot-and-mouth disease virus (fmdv), here ...200919911056
improvement of a recombinant antibody-based serological assay for foot-and-mouth disease virus.differentiating foot-and-mouth disease virus (fmdv) antibodies generated during a natural infection from those due to vaccination (diva) is crucial for proving freedom from disease after an outbreak and allowing resumption of trade in livestock products. the world organisation for animal health (oie) recommends that fmdv vaccines are composed of inactivated virus that has been purified to remove non-structural viral proteins. such purified vaccines primarily induce antibodies to viral structural ...201019913020
structural insights into replication initiation and elongation processes by the fmdv rna-dependent rna polymerase.rna-dependent rna polymerases (rdrps) play central roles in both transcription and viral genome replication. in picornaviruses, these functions are catalyzed by the virally encoded rdrp, termed 3d. polymerase 3d also catalyzes the covalent linkage of ump to a tyrosine on the small protein vpg. uridylylated vpg then serves as a protein primer for the initiation of rna synthesis. seven different crystal structures of foot-and-mouth disease virus (fmdv) 3d catalytic complexes have enhanced our unde ...200919914060
structure-function analysis of mutant rna-dependent rna polymerase complexes with vpg.the replication of the foot-and-mouth disease virus (fmdv) genome is critically dependent upon the activity of a virally encoded rna-dependent rna polymerase (rdrp). in this study, four mutant rdrps of fmdv were isolated from viral quasi-species treated with ribavirin, of which two were single mutants (l123f and t381a) and two were double mutants (t291i/t381i and l123f/f244l). the mutant proteins were expressed in escherichia coli and purified by his-bind resin chromatography. in combination wit ...200919916926
phylogenetic analysis of 3c protease (3c(pro)) coding region of foot-and-mouth disease virus type a.nucleotide sequence analysis of the 3c protease (3c(pro)) region of foot-and-mouth disease virus type a (fmdv-a) isolates from india has revealed incongruous phylogenetic grouping between 3c(pro) and vp1 region possibly due to the genetic recombination or independent evolution of non-structural and structural protein coding regions. similar to the vp1 region, the emerging vp3(59)-deletion group maintained its genetic distinctiveness at 3c(pro) region and was found to be diverging with time. two ...200919941399
cd40-expressing plasmid induces anti-cd40 antibody and enhances immune responses to dna vaccination.various approaches have been used to improve the efficacy of dna vaccination, including the incorporation of molecular adjuvants. because the cd40 ligand-cd40 interaction plays a major role in initiating immune responses, we sought to develop a molecular adjuvant targeting this interaction.201019950201
interferon-induced protection against foot-and-mouth disease virus infection correlates with enhanced tissue-specific innate immune cell infiltration and interferon-stimulated gene expression.previously, we demonstrated that type i interferon (ifn-alpha/beta) or a combination of ifn-alpha/beta and type ii ifn (ifn-gamma) delivered by a replication-defective human adenovirus 5 (ad5) vector protected swine when challenged 1 day later with foot-and-mouth disease virus (fmdv). to gain a more comprehensive understanding of the mechanism of protection induced by ifns, we inoculated groups of six swine with ad5-vectors containing these genes, challenged 1 day later and euthanized 2 animals ...201019955313
currently important animal disease management issues in sub-saharan africa.the present international approach to management of transboundary animal diseases (tads) is based on the assumption that most can be eradicated; consequently, that is the usual objective adopted by international organizations concerned with animal health. however, for sub-saharan africa and southern africa more particularly, eradication of most tads is impossible for the foreseeable future for a variety of technical, financial and logistical reasons. compounding this, the present basis for acces ...200919967938
molecular characterization of sat 2 foot-and-mouth disease virus from post-outbreak slaughtered animals: implications for disease control in uganda, limiting the extent of foot-and-mouth disease (fmd) spread during outbreaks involves short-term measures such as ring vaccination and restrictions of the movement of livestock and their products to and from the affected areas. in this study, the presence of fmd virus rna was investigated in cattle samples 3 months after fmd quarantine measures had been lifted following an outbreak in 2004. oropharyngeal tissue samples were obtained from 12 cattle slaughtered in a small town abattoir i ...201020003615
rna immunization can protect mice against foot-and-mouth disease previous work we have reported the immunization of swine using in vitro-transcribed foot-and-mouth disease virus (fmdv) rna. with the aim of testing whether rna-induced immunization can mediate protection against viral infection, a group of swiss adult mice was inoculated with fmdv infectious transcripts. in most inoculated animals viral rna was detected in serum at 48-72h postinoculation. a group of the rna-inoculated mice (11 out of 19) developed significant titers of neutralizing antibodie ...201020005905
recovery of infectious foot-and-mouth disease virus from full-length genomic cdna clones using an rna polymerase i system.the prototypic foot-and-mouth disease virus (fmdv) was shown more than a century ago to be the first filterable agent capable of causing fmd, and it has served as an important model for studying basic principles of aphthovirus molecular biology. however, the complex structure and antigenic diversity of fmdv have posed a major obstacle to the attempts at manipulating the infectious virus by reverse genetic techniques. here, we report the recovery of infectious fmdv from cdnas based on an efficien ...200920011974
molecular cloning and phylogenetic analysis of integrins alphavbeta1 and alphavbeta6 of one-humped camel (camelus dromedarius).bactrian camels can relatively easily be infected with fmdv, but dromedary camels remain resistant even to high doses of the virus. to understand the different susceptibility between the two camel species from the standpoint of viral receptors, this work reports the sequences of the dromedary camel integrin cdnas encoding alphavbeta1 and alphavbeta6 and compare them to those of other species, especially to bactrian camels. the complete coding sequences for the dromedary camel alphav, beta1 and b ...201020015555
design and optimization of a novel reverse transcription linear-after-the-exponential pcr for the detection of foot-and-mouth disease virus.a novel molecular assay for the detection of foot-and-mouth disease virus (fmdv) was developed using linear-after-the-exponential polymerase chain reaction (late-pcr).201020028437
rt-pcr evaluation for identification and sequence analysis of foot-and-mouth disease serotype o from 2006 to 2007 in punjab, pakistan.fmd clinically positive 250 tissue samples (mouth and hoof epithelium and vesicle swabs, tongue tissue) and 175 secretion samples (milk, saliva, serum, plasma) were evaluated by rt-pcr for the diagnosis of fmd with different pair of universal and serotype-specific primers from 2006 to 2007 in punjab, pakistan. universal primer pair p1/p2 from vp1 gene detected fmd in 182 out of 250 (72.8%) tissues and 92 out of 175 (52.6%) secretion samples, while universal primer 1f/1r from 5'utr region detecte ...201120031216
high expression level of a foot and mouth disease virus epitope in tobacco transplastomic plants.chloroplast transformation has an extraordinary potential for antigen production in plants because of the capacity to accumulate high levels of recombinant proteins and increased biosafety due to maternal plastid inheritance in most crops. in this article, we evaluate tobacco chloroplasts transformation for the production of a highly immunogenic epitope containing amino acid residues 135-160 of the structural protein vp1 of the foot and mouth disease virus (fmdv). to increase the accumulation le ...201020041332
foot-and-mouth disease viral loads in pigs in the early, acute stage of disease.the progress and pathogenesis of foot-and-mouth disease virus (fmdv) was studied in infected pigs by observing the development of clinical signs in two separate experiments. viral loads were determined by real-time quantitative rt-pcr in the liver, spleen, cervical lymph node, mandibular lymph node, retropharyngeal lymph node, soft palate, pharynx, tonsil, tongue and skin (coronary band area). tissue samples were collected from both inoculated and contact-infected pigs at several time points dur ...201020045852
the effect of foot-and-mouth disease (fmd) vaccination on virus transmission and the significance for the field.vaccination against foot-and-mouth disease (fmd) might be one of the control measures used during an fmd epidemic depending on the local epidemiological situation, the status of the country, and the opinion of policy makers. a sound decision on vaccination can be made only if there is sufficient scientific knowledge on the effectiveness of vaccination in eliminating the virus from the population. an important question is whether a single vaccination applied as an emergency vaccine can contribute ...200920046605
a single amino acid substitution in the capsid of foot-and-mouth disease virus can increase acid lability and confer resistance to acid-dependent uncoating inhibition.the acid-dependent disassembly of foot-and-mouth disease virus (fmdv) is required for viral rna release from endosomes to initiate replication. although the fmdv capsid disassembles at acid ph, mutants escaping inhibition by nh(4)cl of endosomal acidification were found to constitute about 10% of the viruses recovered from bhk-21 cells infected with fmdv c-s8c1. for three of these mutants, the degree of nh(4)cl resistance correlated with the sensitivity of the virion to acid-induced inactivation ...201020053737
effect of the extract made from cochinchina momordica seeds on the humoral immune responses of mice to a commercial foot-and-mouth disease vaccine (serotypes o and asia 1).the extract from ecms was investigated for its effect on the humoral immune responses to foot-and-mouth disease vaccination. fifty-six mice were randomly divided into seven groups with eight animals in each. mice in groups 5 to 7 were subcutaneously (s.c.) injected with 0.5 mg dex daily for 4 days to induce immunosuppression. the animals were then orally given ecms (200 microg in 250 microl saline) in groups 3 and 6 or 250 microl saline in group 2, or s.c. injected with ecms (50 microg in 100 mi ...201020055940
phylogenetic analysis of indian serotype asia1 foot-and-mouth-disease virus isolates revealed emergence and reemergence of different genetic lineages.foot-and-mouth-disease virus (fmdv) serotype asia1 causes significant number of disease outbreaks in india. indian asia1 virus isolates were shown to be genetically heterogeneous and of the two lineages (lineage b and lineage c) described in india, lineage c caused majority of the outbreaks. emergence of a novel divergent lineage (lineage d) within lineage c has been described in 2001. in the present report, the complete vp1 genomic region of 41 fmdv asia1 field isolates collected between 2003 a ...201020080365
competition-colonization dynamics in an rna virus.during replication, rna viruses rapidly generate diverse mutant progeny which differ in their ability to kill host cells. we report that the progeny of a single rna viral genome diversified during hundreds of passages in cell culture and self-organized into two genetically distinct subpopulations that exhibited the competition-colonization dynamics previously recognized in many classical ecological systems. viral colonizers alone were more efficient in killing cells than competitors in culture. ...201020080701
possible use of rna isolate from inactivated vaccine for external positive control in reverse transcription-based detection of foot-and-mouth disease virus in bull semen.this study has demonstrated the novel use of inactivated and purified vaccine against fmd virus for detection and analysis. rna isolate was efficiently generated from the vaccine for an external positive control for reverse transcription polymerase chain reaction (rt-pcr) and reverse transcription loop-mediated isothermal amplification (rt-lamp) assays. the target dna fragment sequences from the 2b region and 3d rna polymerase gene of the virus for rt-pcr and rt-lamp respectively were successful ...201020097168
molecular characterization of serotype a foot-and-mouth disease viruses circulating in vietnam in 2009.foot-and-mouth disease (fmd) is a major cause of endemic outbreaks in vietnam in recent years. in this work, six serotype a foot-and-mouth disease viruses (fmdv), collected from endemic outbreaks during january and february of 2009 in four different provinces in vietnam, were genetically characterized for their complete genome sequences. genetic analysis based on the complete viral genome sequence indicated that they were closely related to each other and shared 99.0-99.8% amino acid (aa) identi ...201020097490
development of an immunochromatographic strip for the detection of antibodies against foot-and-mouth disease virus serotype immunochromatographic strip was developed for the serological detection of type o foot-and-mouth disease (fmd) in swine. in the strip, the expressed protein of vp1, the main protective antigen of fmdv, labeled with colloidal gold was used as the detector, the staphylococcal protein a (spa) and swine anti-foot-and-mouth disease virus (fmdv) antibody were blotted on the nitrocellulose membrane for the test and control lines, respectively. 296 swine serum samples were collected to evaluate the c ...201020100515
iga antibody response of swine to foot-and-mouth disease virus infection and vaccination.foot-and-mouth disease virus (fmdv) continues to be a significant economic problem worldwide. control of the disease involves the use of killed-virus vaccines, a control measure developed decades ago. after natural infection, the primary site of replication of fmdv is the pharyngeal area, suggesting that a mucosal immune response is the most effective. humoral immunity to killed-virus vaccination induces antibodies that can prevent the clinical disease but not local infection. determining whethe ...201020107003
loss of plasmacytoid dendritic cell function coincides with lymphopenia and viremia during foot-and-mouth disease virus infection.foot-and-mouth disease virus (fmdv) causes an acute, highly contagious disease of livestock. though fmdv is very sensitive to interferon-alpha (ifn-alpha), ifn-beta, and ifn-gamma, the virus has evolved mechanisms to evade such innate responses. for instance, during acute infection, fmdv suppresses ifn-alpha production by skin and myeloid dendritic cells (dcs). we have previously reported that fmdv infection induces a transient lymphopenia and interruption of t-lymphocyte responses to mitogenic ...201020121400
a bicistronic lentiviral vector based on the 1d/2a sequence of foot-and-mouth disease virus expresses proteins stoichiometrically.classic ires sequences are notorious for exerting biased expression in favor of upstream coding regions when placed into polycistronic vectors. here, we report the development of a bicistronic lentiviral system based on the 1d/2a sequence from the foot-and-mouth disease virus that is able to maintain tightly balanced control of upstream and downstream protein expression for several days at a stoichiometry very closely approaching 1.0. our results suggest that the 1d/2a sequence can be optimized ...201020132845
recombinant dna and protein vaccines for foot-and-mouth disease induce humoral and cellular immune responses in mice.foot-and-mouth disease virus (fmdv) is a small single-stranded rna virus which belongs to the family picornaviridae, genus apthovirus. it is a principal cause of fmd which is highly contagious in livestock. in a wild type virus infection, infected animals usually elicit antibodies against structural and non-structural protein of fmdv. a structural protein, vp1, is involved in neutralization of virus particle, and has both b and t cell epitopes. a rna-dependent rna polymerase, 3d, is highly conse ...200920157614
attenuated salmonella choleraesuis-mediated rnai targeted to conserved regions against foot-and-mouth disease virus in guinea pigs and this study, specific sequences within three genes (3d, vp4 and 2b) of the foot-and-mouth disease virus (fmdv) genome were determined to be effective rnai targets. these sequences are highly conserved among different serotype viruses based on sequence analysis. small interfering rna (sirna)-expressing plasmids (p3d-nt19, p3d-nt56, pvp4-nt19, pvp4-nt65 and p2b-nt25) were constructed to express sirna targeting 3d, vp4 and 2b, respectively. the antiviral potential of these sirna for various fmdv ...201020167192
hsp70 enhances presentation of fmdv antigen to bovine cd4+ t cells in vitro.foot-and-mouth disease virus (fmdv) is the causative agent of a highly contagious acute vesicular disease affecting cloven-hoofed animals, including cattle, sheep and pigs. the current vaccine induces a rapid humoral response, but the duration of the protective antibody response is variable, possibly associated with a variable specific cd4+ t cell response. we investigated the use of heat shock protein 70 (hsp70) as a molecular chaperone to target viral antigen to the major histocompatibility co ...201020167197
quantitative single serum-dilution liquid phase competitive blocking elisa for the assessment of herd immunity and expected protection against foot-and-mouth disease virus in vaccinated cattle.a single serum-dilution liquid phase elisa (slpelisa) was standardized to be used for serological evaluation of herd immunity against foot-and-mouth disease. the absorbance value at a dilution 1:64 of each serum sample was interpolated in a standard curve by plotting the antibody titers of six control sera determined by end point dilution liquid phase elisa (lpelisa), against the absorbance values for the same control sera at 1:64 dilutions. a straight line was obtained by linear regression anal ...201020170683
transgenic mouse model integrating sirna targeting the foot and mouth disease virus.we have constructed 2 small interfering rnas (sirnas) specifically targeting homogenous 3d and 2b1 regions of 7 serotypes of the foot and mouth disease virus (fmdv) and tested the ability of sirnas to inhibit virus replication in baby hamster kidney (bhk-21) cells and suckling mice. in this study, we generated transgenic mouse models integrating short hairpin rna (shrna) targeting microinfected fmdv. when examined at the 7th passage in transgenic mice, the target gene was still found by pcr to b ...201020176056
limiting the impacts of foot and mouth disease in large ruminants in northern lao people's democratic republic by vaccination: a case study.foot and mouth disease (fmd) is the most important global transboundary livestock disease and is endemic in lao people's democratic republic (lao pdr) with outbreaks occurring regularly. lao pdr shares borders with five countries and as a major thoroughfare for transboundary livestock movement, is vulnerable to the social and economic impacts of fmd. the fmd outbreak occurred in january 2009 in the pek district, located in the north-eastern lao pdr province of xieng khuang and involved all 111 v ...201020180923
a novel system for the production of high levels of functional human therapeutic proteins in stable cells with a semliki forest virus noncytopathic vector.semliki forest virus (sfv) vectors lead to high protein expression in mammalian cells, but expression is transient due to vector cytopathic effects, inhibition of host cell proteins and rna-based expression. we have used a noncytopathic sfv mutant (ncsfv) rna vector to generate stable cell lines expressing two human therapeutic proteins: insulin-like growth factor i (igf-i) and cardiotrophin-1 (ct-1). therapeutic genes were fused at the carboxy-terminal end of puromycin n-acetyl-transferase gene ...201020188220
the relationship between cellular immune response to foot-and-mouth disease virus asia 1 and viral persistence in indian cattle (bosindicus).foot-and-mouth disease (fmd), the most contagious animal disease, is associated with persistent viral infection in ruminants, despite the induction of systemic immune response. the present study was performed to decipher the relation between the persistent fmd virus (fmdv) infection and cellular immune response in indian cattle (bosindicus) following experimental inoculation of fmdv asia 1. persistent viral infection (carriers) was detected by antigen capture rt-pcr on the oesophageal-pharyngeal ...201020189208
tailoring the switch from ires-dependent to 5'-end-dependent translation with the rnase p ribozyme.translation initiation driven by internal ribosome entry site (ires) elements is dependent on the structural organization of the ires region. we have previously shown that a structural motif within the foot-and-mouth-disease virus ires is recognized in vitro as substrate for the synechocystis sp. rnase p ribozyme. here we show that this structure-dependent endonuclease recognizes the ires element in cultured cells, leading to inhibition of translation. inhibition of ires activity was dependent o ...201020194518
a recombinant truncated fmdv 3ab protein used to better distinguish between infected and vaccinated distinguish the antibodies induced by foot-and-mouth disease virus (fmdv) infection from those induced by vaccination, a recombinant n-terminal truncated fmdv non-structural protein (nsp) of 3ab, designated as r3ab, was constructed by deleting 80 amino acids displayed about 30% homology to transposase is4 family protein of escherichia coli, expressed in e. coli bl21 (de3) and then purified. the r3ab was majorly expressed in soluble fraction and presented as homogeneous monomers after purifica ...201020197135
marker vaccine potential of a foot-and-mouth disease virus with a partial vp1 g-h loop deletion.previous work in cattle and pigs demonstrated that protection against foot-and-mouth disease (fmd) could be achieved following vaccination with chimeric foot-and-mouth disease virus (fmdv) vaccines, in which the vp1 g-h loop had been substituted with that from another serotype. this indicated that the vp1 g-h loop may not be essential for the protection of natural hosts against fmdv. if this could be substantiated there would be potential to develop fmd marker vaccines, characterised by the abse ...201020199761
the development of oral fluid-based diagnostics and applications in veterinary medicine.the purpose of this review was to discuss the history of the development and implementation of oral fluid diagnostics for infectious diseases of humans and domestic animals. the use of oral fluid for the assessment of health and diagnosis of disease in humans and animals has a surprisingly long history. as early as 1909, pollaci and ceraulo reported sensitive and specific agglutination of 'micrococcus melitensis' (brucella melitensis) by oral fluid from patients diagnosed with malta fever. a 198 ...201020202287
parameterization of the duration of infection stages of serotype o foot-and-mouth disease virus: an analytical review and meta-analysis with application to simulation models.foot-and-mouth disease (fmd) is considered one of the most important infectious diseases of livestock because of the devastating economic consequences that it inflicts in affected regions. the value of critical parameters, such as the duration of the latency or the duration of the infectious periods, which affect the transmission rate of the fmd virus (fmdv), are believed to be influenced by characteristics of the host and the virus. disease control and surveillance strategies, as well as fmd si ...201020205988
characterization of foot-and-mouth disease virus antigen by surface-enhanced laser desorption ionization-time of flight-mass spectrometry in aqueous and oil-emulsion formulations.we have used a novel method, surface-enhanced laser desorption ionization-time of flight-mass spectrometry (seldi-tof-ms), to characterize foot-and-mouth disease virus (fmdv) vaccine antigens. using specific capture with fmdv binding recombinant antibody fragments and tryptic digestion of fmdv antigens the spectral peaks representing the fmdv structural proteins vp1, vp2, vp3 and vp4 were identified. vp1 existed as 2 variants differing by 0.2kda and vp4 as 8 variants differing by 14-17da. such h ...201020206283
[morphological observation of bovine kidney (mdbk) cells effected by foot-and-mouth disease virus l(pro)].in order to explore the morphological changes of bovine kidney (mdbk) cells induced by foot-and-mouth disease virus (fmdv) l protease, we induced the expression of fmdv l protease in bovine kidney cells (mdbk) artificially. all work is carried out on the basis of a stable mdbk cell line inducibly expresses the lab gene under the control of tetracycline. we use cell morphology, hoechst 33258 staining, ao-eb staining, and dna ladder abstraction to research the morphological changes of mdbk cells. ...200920222457
[rescue of bovine asia 1 serotype foot-and-mouth disease virus from a full-length cdna clone].after sequencing the asia 1 foot-and-mouth disease virus (fmdv) (as01 strain), we amplified the two fragments covering the whole genome by overlapping pcr and long pcr. the 5' fragment was 1.8 kb in length including 15cs, and the 3' fragment was 6.7 kb in length. the two fragments were cloned into the pbluescript sk vector to construct recombinant plasmid pbsas carrying the full-length cdna of fmdv as01 strain. the rna transcript was synthesized in vitro using t7 polymerase and transfected into ...200920222458
[not available]. 194720244008
[not available]. 194720244009
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