Utilize este identificador para referenciar este registo: http://hdl.handle.net/10400.21/2159
Título: A new cloning system based on the OprI lipoprotein for the production of recombinant bacterial cell wall-derived immunogenic formulations
Autor: Basto, Afonso P.
Piedade, João
Ramalho, Ruben
Alves, Susana
Soares, Helena
Cornelis, Pierre
Martins, Carlos
Leitão, Alexandre
Palavras-chave: African swine fever virus/genetics
Animals
Antigens, viral/genetics
Bacterial proteins/biosynthesis
Bacterial proteins/genetics
Bacterial proteins/immunology
Base sequence
Cells, cultured
Cloning, molecular/methods
Dendritic cells/immunology
Escherichia coli/genetics
Escherichia coli/metabolism
Genetic vectors/genetics
Lipoproteins/biosynthesis
Lipoproteins/genetics
Lipoproteins/immunology
Macrophages/immunology
Mice
Molecular sequence data
Pseudomonas aeruginosa/genetics
Recombinant fusion proteins/biosynthesis
Recombinant fusion proteins/genetics
Recombinant fusion proteins/immunology
Tumor necrosis factor-alpha/metabolism
Data: Jan-2012
Editora: Elsevier
Citação: Basto AP, Piedade J, Ramalho R, Alves S, Soares H, Cornelis P, et al. A new cloning system based on the OprI lipoprotein for the production of recombinant bacterial cell wall-derived immunogenic formulations. J Biotechnol. 2012;157(1):50-63.
Resumo: The conjugation of antigens with ligands of pattern recognition receptors (PRR) is emerging as a promising strategy for the modulation of specific immunity. Here, we describe a new Escherichia coli system for the cloning and expression of heterologous antigens in fusion with the OprI lipoprotein, a TLR ligand from the Pseudomonas aeruginosa outer membrane (OM). Analysis of the OprI expressed by this system reveals a triacylated lipid moiety mainly composed by palmitic acid residues. By offering a tight regulation of expression and allowing for antigen purification by metal affinity chromatography, the new system circumvents the major drawbacks of former versions. In addition, the anchoring of OprI to the OM of the host cell is further explored for the production of novel recombinant bacterial cell wall-derived formulations (OM fragments and OM vesicles) with distinct potential for PRR activation. As an example, the African swine fever virus ORF A104R was cloned and the recombinant antigen was obtained in the three formulations. Overall, our results validate a new system suitable for the production of immunogenic formulations that can be used for the development of experimental vaccines and for studies on the modulation of acquired immunity.
Peer review: yes
URI: http://hdl.handle.net/10400.21/2159
ISSN: 0168-1656
Versão do Editor: http://www.sciencedirect.com/science/article/pii/S0168165611006407
Aparece nas colecções:ESTeSL - Artigos

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