A new plasmid vector for DNA delivery using lactococci
| Autor: | Vasco Azevedo, François Lefèvre, Jean-Marc Chatel, Anderson Miyoshi, Sylvia Innocentin, Valeria Guimarães, Philippe Langella |
|---|---|
| Přispěvatelé: | Instituto de Ciencias Biologicas, Universidade Federal de Minas Gerais (UFMG), Unité de recherche d'Écologie et Physiologie du Système Digestif (UEPSD), Institut National de la Recherche Agronomique (INRA), Unité de Recherche Immuno-Allergie Alimentaire, Unité de recherche Virologie et Immunologie Moléculaires (VIM (UR 0892)) |
| Rok vydání: | 2008 |
| Předmět: |
Cloning
0303 health sciences biology 030306 microbiology Genetic enhancement [SDV]Life Sciences [q-bio] Research Lactococcus lactis Immunology Transfection biology.organism_classification Molecular biology Green fluorescent protein DNA vaccination 03 medical and health sciences Plasmid Antigen Immunology and Allergy Molecular Medicine 030304 developmental biology Biotechnology |
| Zdroj: | Genetic Vaccines and Therapy Genetic Vaccines and Therapy, BioMed Central, 2009, 7 (4), 7 p. ⟨10.1186/1479-0556-7-4⟩ Genetic Vaccines and Therapy 4 (7), 7 p.. (2009) |
| ISSN: | 1479-0556 |
| DOI: | 10.1186/1479-0556-7-4⟩ |
| Popis: | Background The use of food-grade lactococci as bacterial carriers to DNA delivery into epithelial cells is a new strategy to develop live oral DNA vaccine. Our goal was to develop a new plasmid, named pValac, for antigen delivery for use in lactococci. The pValac plasmid was constructed by the fusion of: i) a eukaryotic region, allowing the cloning of an antigen of interest under the control of the pCMV eukaryotic promoter to be expressed by a host cell and ii) a prokaryotic region allowing replication and selection of bacteria. In order to evaluate pValac functionality, the gfp ORF was cloned into pValac (pValac:gfp) and was analysed by transfection in PK15 cells. The applicability of pValac was demonstrated by invasiveness assays of Lactococcus lactis inlA+ strains harbouring pValac:gfp into Caco-2 cells. Results After transfection with pValac:gfp, we observed GFP expression in PK15 cells. L. lactis inlA+ were able to invade Caco-2 cells and delivered a functional expression cassette (pCMV:gfp) into epithelial cells. Conclusion We showed the potential of an invasive L. lactis harbouring pValac to DNA delivery and subsequent triggering DNA expression by epithelial cells. Further work will be to examine whether these strains are able to deliver DNA in intestinal cells in vivo. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full text from SpringerLink