scholarly journals A Novel Plasmid DNA-Based Foot and Mouth Disease Virus Minigenome for Intracytoplasmic mRNA Production

Viruses ◽  
2021 ◽  
Vol 13 (6) ◽  
pp. 1047
Author(s):  
Ploypailin Semkum ◽  
Challika Kaewborisuth ◽  
Nattarat Thangthamniyom ◽  
Sirin Theerawatanasirikul ◽  
Chalermpol Lekcharoensuk ◽  
...  
Keyword(s):  
Reverse Genetics ◽  
Antiviral Drug ◽  
Foot And Mouth Disease ◽  
Mouth Disease ◽  
Foreign Gene ◽  
Efficient Production ◽  
Rna Transfection ◽  
Mouth Disease Virus ◽  
Foot And Mouth ◽  
Minigenome System

Picornaviruses are non-enveloped, single-stranded RNA viruses that cause highly contagious diseases, such as polio and hand, foot-and-mouth disease (HFMD) in human, and foot-and-mouth disease (FMD) in animals. Reverse genetics and minigenome of picornaviruses mainly depend on in vitro transcription and RNA transfection; however, this approach is inefficient due to the rapid degradation of RNA template. Although DNA-based reverse genetics systems driven by mammalian RNA polymerase I and/or II promoters display the advantage of rescuing the engineered FMDV, the enzymatic functions are restricted in the nuclear compartment. To overcome these limitations, we successfully established a novel DNA-based vector, namely pKLS3, an FMDV minigenome containing the minimum cis-acting elements of FMDV essential for intracytoplasmic transcription and translation of a foreign gene. A combination of pKLS3 minigenome and the helper plasmids yielded the efficient production of uncapped-green florescent protein (GFP) mRNA visualized in the transfected cells. We have demonstrated the application of the pKLS3 for cell-based antiviral drug screening. Not only is the DNA-based FMDV minigenome system useful for the FMDV research and development but it could be implemented for generating other picornavirus minigenomes. Additionally, the prospective applications of this viral minigenome system as a vector for DNA and mRNA vaccines are also discussed.

scholarly journals An infectious recombinant foot-and-mouth disease virus expressing a fluorescent marker protein

2013 ◽  
Vol 94 (7) ◽  
pp. 1517-1527 ◽  
Author(s):  
Julian Seago ◽  
Nicholas Juleff ◽  
Katy Moffat ◽  
Stephen Berryman ◽  
John M. Christie ◽  
...  
Keyword(s):  
Disease Virus ◽  
Reverse Genetics ◽  
Foot And Mouth Disease ◽  
Mouth Disease ◽  
Marker Protein ◽  
Fluorescent Marker ◽  
Fmdv Infection ◽  
Animal Pathogens ◽  
Mouth Disease Virus ◽  
Foot And Mouth

Foot-and-mouth disease virus (FMDV) is one of the most extensively studied animal pathogens because it remains a major threat to livestock economies worldwide. However, the dynamics of FMDV infection are still poorly understood. The application of reverse genetics provides the opportunity to generate molecular tools to further dissect the FMDV life cycle. Here, we have used reverse genetics to determine the capsid packaging limitations for a selected insertion site in the FMDV genome. We show that exogenous RNA up to a defined length can be stably introduced into the FMDV genome, whereas larger insertions are excised by recombination events. This led us to construct a recombinant FMDV expressing the fluorescent marker protein, termed iLOV. Characterization of infectious iLOV-FMDV showed the virus has a plaque morphology and rate of growth similar to the parental virus. In addition, we show that cells infected with iLOV-FMDV are easily differentiated by flow cytometry using the inherent fluorescence of iLOV and that cells infected with iLOV-FMDV can be monitored in real-time with fluorescence microscopy. iLOV-FMDV therefore offers a unique tool to characterize FMDV infection in vitro, and its applications for in vivo studies are discussed.

scholarly journals The broad-spectrum antiviral drug arbidol inhibits foot-and-mouth disease virus replication

2019 ◽  
Vol 1 (1A) ◽  
Author(s):  
Stephen Polyak ◽  
Morgan Herod ◽  
Oluwapelumi Adeyemi ◽  
Joseph Ward ◽  
Toby Tuthill ◽  
...  
Keyword(s):  
Disease Virus ◽  
Virus Replication ◽  
Broad Spectrum ◽  
Antiviral Drug ◽  
Foot And Mouth Disease ◽  
Mouth Disease ◽  
Mouth Disease Virus ◽  
Foot And Mouth

scholarly journals Structural and Mutagenic Analysis of Foot-and-Mouth Disease Virus 3C Protease Reveals the Role of the β-Ribbon in Proteolysis

2006 ◽  
Vol 81 (1) ◽  
pp. 115-124 ◽  
Author(s):  
Trevor R. Sweeney ◽  
Núria Roqué-Rosell ◽  
James R. Birtley ◽  
Robin J. Leatherbarrow ◽  
Stephen Curry
Keyword(s):  
Catalytic Activity ◽  
Disease Virus ◽  
Serine Proteases ◽  
Antiviral Drug ◽  
Foot And Mouth Disease ◽  
Mouth Disease ◽  
3C Protease ◽  
Mouth Disease Virus ◽  
Foot And Mouth

ABSTRACT The 3C protease (3Cpro) from foot-and-mouth disease virus (FMDV), the causative agent of a widespread and economically devastating disease of domestic livestock, is a potential target for antiviral drug design. We have determined the structure of a new crystal form of FMDV 3Cpro, a chymotrypsin-like cysteine protease, which reveals features that are important for catalytic activity. In particular, we show that a surface loop which was disordered in previous structures adopts a β-ribbon structure that is conformationally similar to equivalent regions on other picornaviral 3C proteases and some serine proteases. This β-ribbon folds over the peptide binding cleft and clearly contributes to substrate recognition. Replacement of Cys142 at the tip of the β-ribbon with different amino acids has a significant impact on enzyme activity and shows that higher activity is obtained with more hydrophobic side chains. Comparison of the structure of FMDV 3Cpro with homologous enzyme-peptide complexes suggests that this correlation arises because the side chain of Cys142 contacts the hydrophobic portions of the P2 and P4 residues in the peptide substrate. Collectively, these findings provide compelling evidence for the role of the β-ribbon in catalytic activity and provide valuable insights for the design of FMDV 3Cpro inhibitors.

scholarly journals The broad-spectrum antiviral drug arbidol inhibits foot-and-mouth disease virus genome replication

2019 ◽  
Vol 100 (9) ◽  
pp. 1293-1302 ◽  
Author(s):  
Morgan R. Herod ◽  
Oluwapelumi O. Adeyemi ◽  
Joseph Ward ◽  
Kirsten Bentley ◽  
Mark Harris ◽  
...  
Keyword(s):  
Disease Virus ◽  
Broad Spectrum ◽  
Antiviral Drug ◽  
Foot And Mouth Disease ◽  
Virus Genome ◽  
Mouth Disease ◽  
Genome Replication ◽  
Mouth Disease Virus ◽  
Foot And Mouth

scholarly journals In Vitro and in Vivo Antiviral Activity of Mizoribine Against Foot-And-Mouth Disease Virus

Molecules ◽  
2019 ◽  
Vol 24 (9) ◽  
pp. 1723 ◽  
Author(s):  
Shi-Fang Li ◽  
Mei-Jiao Gong ◽  
Yue-Feng Sun ◽  
Jun-Jun Shao ◽  
Yong-Guang Zhang ◽  
...  
Keyword(s):  
Antiviral Activity ◽  
Early Stage ◽  
Antiviral Agents ◽  
Antiviral Drug ◽  
Foot And Mouth Disease ◽  
Mouth Disease ◽  
Mouth Disease Virus ◽  
Foot And Mouth

Foot-and-mouth disease (FMD) is a highly contagious viral disease of cloven-hoofed animals, which has significant economic consequences in affected countries. As the currently available vaccines against FMD provide no protection until 4–7 days post-vaccination, the only alternative method to control the spread of FMD virus (FMDV) during outbreaks is the application of antiviral agents. Hence, it is important to identify effective antiviral agents against FMDV infection. In this study, we found that mizoribine has potent antiviral activity against FMDV replication in IBRS-2 cells. A time-of-drug-addition assay demonstrated that mizoribine functions at the early stage of replication. Moreover, mizoribine also showed antiviral effect on FMDV in vivo. In summary, these results revealed that mizoribine could be a potential antiviral drug against FMDV.

scholarly journals Identification of the largest non-essential regions of the C-terminal portion in 3A protein of foot-and-mouth disease virus for replication in cell culture

2020 ◽  
Vol 17 (1) ◽  
Author(s):  
Pinghua Li ◽  
Xueqing Ma ◽  
Xingwen Bai ◽  
Pu Sun ◽  
Hong Yuan ◽  
...  
Keyword(s):  
Cell Culture ◽  
Disease Virus ◽  
Fluorescent Protein ◽  
Foot And Mouth Disease ◽  
Mouth Disease ◽  
Nucleotide Sequencing ◽  
Foreign Gene ◽  
Terminal Portion ◽  
Mouth Disease Virus ◽  
Foot And Mouth

Abstract Background Recent study has shown that the C-terminal portion of 3A (amino acids (aa) 81–153) is not essential for foot-and-mouth disease virus replication in cell culture, however, the complete C-terminal portion (aa 77–153) of 3A is highly variable and prone to occur deletions and mutations, therefore, we presume that this region plays a very limited role and probablely is completely nonessential for virus viability. Methods In this study, to identify the largest non-essential region of the C-terminal portion in 3A for FMDV viability, several deletions containing aa 80–153, 77–153 and 76–153 of 3A protein were introduced into an FMDV full-length infectious cDNA clone pOFS by the overlapping extension PCR. Additionally, to explore the importance of the highly conserved residue 76 L of 3A for the FMDV of Cathay topotype, two mutants containing 3A L76I and 3A L76V were generated based on the 3A deletion mutant by point mutation. We also introduced the enhanced green fluorescent protein (eGFP) into one of the 3A deletion mutants by the extension PCR to investigate the genetic flexibility of 3A to express foreign genes. All linearized full plasmids were transfected into BSR/T7 cells to rescue infectious foot-and-mouth disease viruses. The rescused viruses were analyzed by RT-PCR, nucleotide sequencing, immunofluorescence assay and western blot and were characterized by plaque assays and one-step growth kinetics. Results The results demonstrated that the deletion of aa 80–153 and aa 77–153 and the substitutions of 3A L76I and 3A L76V did not affect the production of infectious virus, while the fusion of the eGFP gene to the C-terminus of 3A resulted in nonviable FMDV. Conclusions Our results firstly reported that the aa 77–153 rather than aa 81–153 of 3A protein was dispensable for FMDV replication in cell culture. This study is of great significance for development of FMD marker vaccine and foreign gene expression in the future.

Biophysical Measurement of Molecular Weight of Foot-and-Mouth Disease RNA Fragments

1974 ◽  
Vol 32 ◽  
pp. 262-263
Author(s):  
Sydney S. Breese ◽  
Howard L. Bachrach
Keyword(s):  
Chemical Properties ◽  
Foot And Mouth Disease ◽  
Mouth Disease ◽  
Distilled Water ◽  
Bovine Plasma ◽  
Mouth Disease Virus ◽  
Foot And Mouth ◽  
Carbon Coated ◽  
Rna Fragments ◽  
Physical And Chemical

Continuing studies on the physical and chemical properties of foot-and-mouth disease virus (FMDV) have included electron microscopy of RNA strands released when highly purified virus (1) was dialyzed against demlneralized distilled water. The RNA strands were dried on formvar-carbon coated electron microscope screens pretreated with 0.1% bovine plasma albumin in distilled water. At this low salt concentration the RNA strands were extended and were stained with 1% phosphotungstic acid. Random dispersions of strands were recorded on electron micrographs, enlarged to 30,000 or 40,000 X and the lengths measured with a map-measuring wheel. Figure 1 is a typical micrograph and Fig. 2 shows the distributions of strand lengths for the three major types of FMDV (A119 of 6/9/72; C3-Rezende of 1/5/73; and O1-Brugge of 8/24/73.

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