Transient expression of Human papillomavirus type 16 L1 protein in Nicotiana benthamiana using an infectious tobamovirus vector

Arvind Varsani, Anna Lise Williamson, Debbie Stewart, Edward P. Rybicki

Research output: Contribution to journalArticle

52 Scopus citations

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 μg/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.

Original languageEnglish (US)
Pages (from-to)91-96
Number of pages6
JournalVirus research
Volume120
Issue number1-2
DOIs
StatePublished - Sep 1 2006
Externally publishedYes

Keywords

  • HPV-16
  • Papillomavirus
  • Plant expression
  • TMV vector
  • VLP
  • Vaccine

ASJC Scopus subject areas

  • Cancer Research
  • Virology
  • Infectious Diseases

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