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Publication Details
AFRICAN RESEARCH NEXUS
SHINING A SPOTLIGHT ON AFRICAN RESEARCH
Expression of HPV-11 L1 protein in transgenic Arabidopsis thaliana and Nicotiana tabacum
BMC Biotechnology, Volume 7, Article 56, Year 2007
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Description
Background: We have investigated the possibility and feasibility of producing the HPV-11 L1 major capsid protein in transgenic Arabidopsis thaliana ecotype Columbia and Nicotiana tabacum cv. Xanthi as potential sources for an inexpensive subunit vaccine. Results: Transformation of plants was only achieved with the HPV-11 L1 gene with the C-terminal nuclear localization signal (NLS-) encoding region removed, and not with the full-length gene. The HPV-11 L1 NLS- gene was stably integrated and inherited through several generations of transgenic plants. Plant-derived HPV-11 L1 protein was capable of assembling into virus-like particles (VLPs), although resulting particles displayed a pleomorphic phenotype. Neutralising monoclonal antibodies binding both surface-linear and conformation-specific epitopes bound the A. thaliana-derived particles and - to a lesser degree - the N. tabacum-derived particles, suggesting that plant-derived and insect cell-derived VLPs displayed similar antigenic properties. Yields of up to 12 μg/g of HPV-11 L1 NLS- protein were harvested from transgenic A. thaliana plants, and 2 μg/g from N. tabacum plants - a significant increase over previous efforts. Immunization of New Zealand white rabbits with ∼50 μg of plant-derived HPV-11 L1 NLS- protein induced an antibody response that predominantly recognized insect cell-produced HPV-11 L1 NLS- and not NLS+ VLPs. Evaluation of the same sera concluded that none of them were able to neutralise pseudovirion in vitro. Conclusion: We expressed the wild-type HPV-11 L1 NLS- gene in two different plant species and increased yields of HPV-11 L1 protein by between 500 and 1000-fold compared to previous reports. Inoculation of rabbits with extracts from both plant types resulted in a weak immune response, and antisera neither reacted with native HPV-11 L1 VLPs, nor did they neutralise HPV-11 pseudovirion infectivity. This has important and potentially negative implications for the production of HPV-11 vaccines in plants. © 2007 Kohl et al; licensee BioMed Central Ltd.
Authors & Co-Authors
Kohl, Thomas O.
South Africa, Cape Town
Faculty of Health Sciences
South Africa, Cape Town
University of Cape Town
Hitzeroth, I. I.
South Africa, Cape Town
Faculty of Health Sciences
South Africa, Cape Town
University of Cape Town
Christensen, Neil D.
United States, Hershey
Penn State Cancer Institute
Rybicki, Edward P.
South Africa, Cape Town
Faculty of Health Sciences
South Africa, Cape Town
University of Cape Town
Statistics
Citations: 56
Authors: 4
Affiliations: 3
Identifiers
Doi:
10.1186/1472-6750-7-56
Research Areas
Cancer
Genetics And Genomics
Maternal And Child Health