Abstract
A Tobacco mosaic virus (TMV)-derived vector was used to express a native Human papillomavirus type 16 (HPV-16) L1 gene in Nicotiana benthamiana by means of infectious in vitro RNA transcripts inoculated onto N. benthamiana plants. HPV-16 L1 protein expression was quantitated by enzyme-linked immunosorbent assays (ELISA) after concentration of the plant extract. We estimated that the L1 product yield was 20-37 microg/kg of fresh leaf material. The L1 protein in the concentrated extract was antigenically characterised using the neutralising and conformation-specific Mabs H16:V5 and H16:E70, which bound to the plant-produced protein. Particles observed by transmission electron microscopy were mainly capsomers but virus-like particles (VLPs) similar to those produced in other systems were also present. Immunisation of rabbits with the concentrated plant extract induced a weak immune response. This is the first report of the successful expression of an HPV L1 gene in plants using a plant virus vector.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Antibodies, Monoclonal / immunology
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Antibodies, Viral / immunology
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Antibody Specificity
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Antigens, Viral / genetics
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Antigens, Viral / immunology
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Antigens, Viral / isolation & purification
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Antigens, Viral / metabolism*
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Capsid Proteins / genetics
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Capsid Proteins / immunology
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Capsid Proteins / isolation & purification
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Capsid Proteins / metabolism*
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Enzyme-Linked Immunosorbent Assay
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Genetic Vectors
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Neutralization Tests
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Nicotiana / metabolism*
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Oncogene Proteins, Viral / genetics
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Oncogene Proteins, Viral / immunology
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Oncogene Proteins, Viral / isolation & purification
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Oncogene Proteins, Viral / metabolism*
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Papillomaviridae / immunology
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Plant Leaves / metabolism
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Recombinant Proteins / biosynthesis
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Recombinant Proteins / immunology
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Recombinant Proteins / isolation & purification
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Tobacco Mosaic Virus / genetics*
Substances
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Antibodies, Monoclonal
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Antibodies, Viral
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Antigens, Viral
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Capsid Proteins
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Oncogene Proteins, Viral
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Recombinant Proteins
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L1 protein, Human papillomavirus type 16