부산대학교 도서관

To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/j.virusres.2006.01.022 Byline: Arvind Varsani (b), Anna-Lise Williamson (a)(c), Debbie Stewart (a), Edward P. Rybicki (a)(b) Keywords: Papillomavirus; HPV-16; VLP; Vaccine; Plant expression; TMV vector 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[mu]g/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. Author Affiliation: (a) Institute of Infectious Disease and Molecular Medicine, University of Cape Town, Rondebosch, Cape Town 7701, South Africa (b) Department of Molecular and Cell Biology, University of Cape Town, Rondebosch, Cape Town 7701, South Africa (c) National Health Laboratory Services, Groote Schuur Hospital, PO Observatory, Cape Town 7925, South Africa Article History: Received 9 September 2005; Revised 23 January 2006; Accepted 23 January 2006