Transient expression of Human papillomavirus type 16 L1 protein in Nicotiana benthamiana using an infectious tobamovirus vector.
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.
Varsani
Arvind
A
Department of Molecular and Cell Biology, University of Cape Town, Rondebosch, Cape Town 7701, South Africa. arwind@science.uct.ac.za
Williamson
Anna-Lise
AL
Stewart
Debbie
D
Rybicki
Edward P
EP
D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral / Q000276:immunology; D000918:Antibody Specificity; D000956:Antigens, Viral / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D036022:Capsid Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D005822:Genetic Vectors; D009500:Neutralization Tests; D009856:Oncogene Proteins, Viral / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D027383:Papillomaviridae / Q000276:immunology; D018515:Plant Leaves / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000276:immunology / Q000302:isolation & purification; D014026:Nicotiana / Q000378:metabolism*; D014027:Tobacco Mosaic Virus / Q000235:genetics*
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000956:Antigens, Viral; D036022:Capsid Proteins; D009856:Oncogene Proteins, Viral; D011994:Recombinant Proteins; C063317:L1 protein, Human papillomavirus type 16