Abstract
For effective control of foot-and-mouth disease (FMD), the development of rapid diagnostic systems and vaccines are required against its etiological agent, FMD virus (FMDV). To accomplish this, efficient large-scale expression of the FMDV VP1 protein, with high solubility, needs to be optimized. We attempted to produce high levels of a serotype O FMDV VP1 epitope in Escherichia coli. We identified the subtype-independent serotype O FMDV VP1 epitope sequence and used it to construct a glutathione S-transferase (GST) fusion protein. For efficient production of the FMDV VP1 epitope fused to GST (VP1e-GST), four E. coli strains and three temperatures were examined. The conditions yielding the greatest level of VP1e-GST with highest solubility were achieved with E. coli BL21(DE3) at 25 °C. For high-level production, fed-batch cultures were conducted in 5-l bioreactors. When cells were induced at a high density and complex feeding solutions were supplied, approximately 11 g of VP1e-GST was obtained from a 2.9-l culture. Following purification, the VP1 epitope was used to immunize rabbits, and we confirmed that it induced an immune response.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Animals
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Antibodies, Viral / immunology
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Base Sequence
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Batch Cell Culture Techniques
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Bioreactors
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Capsid Proteins / biosynthesis
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Capsid Proteins / chemistry
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Capsid Proteins / genetics*
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Capsid Proteins / immunology*
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Epitopes / chemistry
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Epitopes / genetics*
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Epitopes / immunology
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Epitopes / metabolism*
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Escherichia coli / genetics*
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Escherichia coli / metabolism
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Foot-and-Mouth Disease / immunology
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Foot-and-Mouth Disease / prevention & control
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Foot-and-Mouth Disease / virology
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Foot-and-Mouth Disease Virus / chemistry
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Foot-and-Mouth Disease Virus / classification
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Foot-and-Mouth Disease Virus / genetics*
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Foot-and-Mouth Disease Virus / immunology*
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Glutathione Transferase / genetics
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Glutathione Transferase / immunology
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Glutathione Transferase / metabolism
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Immunization
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Molecular Sequence Data
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Rabbits
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Recombinant Fusion Proteins / biosynthesis
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Recombinant Fusion Proteins / chemistry
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / isolation & purification
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Reproducibility of Results
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Serotyping
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Viral Vaccines / biosynthesis
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Viral Vaccines / chemistry
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Viral Vaccines / genetics
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Viral Vaccines / immunology
Substances
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Antibodies, Viral
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Capsid Proteins
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Epitopes
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Recombinant Fusion Proteins
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VP1 protein, Foot-and-mouth disease virus
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Viral Vaccines
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Glutathione Transferase