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Characterization of a defensin from the oyster Crassostrea gigas - Recombinant production, folding, solution structure, antimicrobial activities, and gene expression ArchiMer
Gueguen, Yannick; Herpin, Amaury; Aumelas, André; Garnier, Julien; Fievet, Julie; Escoubas, Jean-michel; Bulet, Philippe; Gonzalez, Marcelo; Lelong, Christophe; Favrel, Pascal; Bachere, Evelyne.
In invertebrates, defensins were found in arthropods and in the mussels. Here, we report for the first time the identification and characterization of a defensin (Cg-Def) from an oyster. Cg-def mRNA was isolated from Crassostrea gigas mantle using an expressed sequence tag approach. To gain insight into potential roles of Cg-Def in oyster immunity, we produced the recombinant peptide in Escherichia coli, characterized its antimicrobial activities, determined its solution structure by NMR spectroscopy, and quantified its gene expression in vivo following bacterial challenge of oysters. Recombinant Cg-Def was active in vitro against Gram-positive bacteria but showed no or limited activities against Gram-negative bacteria and fungi. The activity of Cg-Def was...
Tipo: Text Palavras-chave: Crassostrea gigas; Genomic library; Microbiology; Defensin; Oyster.
Ano: 2006 URL: http://archimer.ifremer.fr/doc/2006/publication-2127.pdf
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Cloning and expression analysis of allograft inflammatory factor type 1 in coelomocytes of antarctic sea urchin (Sterechinus neumayeri) ArchiMer
Ovando, Fernanda; Gimpel, Carla; Cardenas, Constanza; Machado Cunha Da Silva, Jose Roberto; De Lorgeril, Julien; Gonzalez, Marcelo.
We have cloned and characterized for the first time an allograft inflammatory factor 1 (Sn-AIF-1) from the Antarctic sea urchin. We report the cloning of Sn-AIF-1 cDNA and the characterization of its expression in coelomocytes after a bacterial challenge. The cDNA Sn-AIF-1 has a size of 608 bp and encodes a polypeptide of 151 aa. The deduced amino acid sequence has a putative size of 17.430 Da, an isoelectric point of 4.92, and shows 2 elongation factor handlike motifs that normally bind calcium ions. BLAST analysis revealed close matches with other known AIF-1. The deduced amino acid sequence of Sn-AIF-1 showed high homology with AIF-1 in vertebrates such as fish, mice, and humans; and in the case of invertebrates, the major degree of identity (55%) was...
Tipo: Text Palavras-chave: Antarctica; Sea urchin; Sterechinus neumayeri; Coelomocytes; Gene expression; AIF-1.
Ano: 2012 URL: http://archimer.ifremer.fr/doc/00098/20955/18572.pdf
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Etude de la réponse immunitaire chez l'huître Crassostrea gigas : caractérisation et fonctions de protéines de reconnaissance aux LPS et d'effecteurs antimicrobiens ArchiMer
Gonzalez, Marcelo.
Research on the immune response of oysters is motivated by the problems of infectious diseases potentially found in the farms. Over the course of the last few years, the development of genomic approaches has allowed us to make headway on characterising genes involved in the cup oyster's (Crassostrea gigas) innate immunity, particularly thanks to various programmes for analysing transcriptome, of type EST (Express Sequence Tag) and subtractive banks. Within the context of this work, over the course of which we have used the data from genomic research and we have also developed biochemical approaches, we were particularly interested in studying the proteins involved in reconnaissance reactions to LPS as well as to antimicrobial effectors. In an earlier part,...
Tipo: Text Palavras-chave: Defense peptides; Polysaccharides; LPS; Hemocytes; Immune response; Crassostrea gigas; Shellfish culture; LPS; Défensine; Peptides antimicrobiens; LBP; BPI; Superoxide dismutase; Hémocytes; Crassostrea gigas; Huître; Mollusque; Immunité.
Ano: 2005 URL: http://archimer.ifremer.fr/doc/2005/these-2098.pdf
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Evidence in oyster of a plasma extracellular superoxide dismutase which binds LPS ArchiMer
Gonzalez, Marcelo; Romestand, Bernard; Fievet, Julie; Huvet, Arnaud; Lebart, M; Gueguen, Yannick; Bachere, Evelyne.
We have characterized in the oyster Crassostrea gigas an extracellular superoxide dismutase (Cg-EcSOD) which appears to bind lipopolysaccharides (LPS). The protein has been purified from the oyster plasma and identified as a Cu/ZnSOD according to its N-terminal sequencing and biological activity. Cg-EcSOD expression and synthesis are restricted to hemocytes as revealed by in situ hybridization and immunocytochemistry. Cg-EcSOD-expressing hemocytes were seen in blood circulation, in connective tissues, and closely associated to endothelium blood vessels. Cg-EcSOD presents in its amino acid sequence a LPS-binding motif found in the endotoxin receptor CD14 and we show that the protein displays an affinity to Escherichia coli bacteria and with LPS and Lipid A....
Tipo: Text Palavras-chave: Beta integrin; Oxidative burst; Hemocyte; Crassostrea gigas; Invertebrate; Mollusc bivalve.
Ano: 2005 URL: http://archimer.ifremer.fr/doc/2005/publication-3583.pdf
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Evidence of a bactericidal permeability increasing protein in an invertebrate, the Crassostrea gigas Cg-BPI ArchiMer
Gonzalez, Marcelo; Gueguen, Yannick; Destoumieux Garzon, Delphine; Romestand, Bernard; Fievet, Julie; Pugniere, M; Roquet, F; Escoubas, Jean-michel; Vandenbulcke, F; Levy, O; Saune, Laure; Bulet, P; Bachere, Evelyne.
A cDNA sequence with homologies to members of the LPS-binding protein and bactericidal/permeability-increasing protein (BPI) family was identified in the oyster Crassostrea gigas. The recombinant protein was found to bind LIPS, to display bactericidal activity against Escherichia coli, and to increase the permeability of the bacterial cytoplasmic membrane. This indicated that it is a BPI rather than an LPS-binding protein. By in situ hybridization, the expression of the C gigas BPI (Cg-bpi) was found to be induced in hemocytes after oyster bacterial challenge and to be constitutive in various epithelia of unchallenged oysters. Thus, Cg-bpi transcripts were detected in the epithelial cells of tissues/organs in contact with the external environment (mantle,...
Tipo: Text Palavras-chave: Oyster innate immunity; Mollusk; Hemocyte; Epithelia; Antimicrobial.
Ano: 2007 URL: http://archimer.ifremer.fr/doc/2007/publication-3564.pdf
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Molecular characterization of two isoforms of defensin from hemocytes of the oyster Crassostrea gigas ArchiMer
Gonzalez, Marcelo; Gueguen, Yannick; Desserre, Geraldine; De Lorgeril, Julien; Romestand, Bernard; Bachere, Evelyne.
Antimicrobial peptides (AMPs) are important components of the host innate immune response against microbial invasion. We previously characterized the first AMP from an oyster, a defensin, that was shown to be continuously expressed in the mantle of Crassostrea gigas. In this study, we report the cDNA cloning of two new isoforms of the defensin AMP family (Cg-defh1 and Cg-defh2) from the hemocytes of the oyster. The deduced amino acid sequences reveal two peptides of 73 amino acid residues with a mature portion consisting of 43 amino acid residues. Cy-Defh1 and CyDefh2 share 86% amino acid identity and belong to the "arthropod-molluscs defensin family". qRT-PCR analyses indicate that Cg-defh2 is continuously expressed in the hemocytes of C. gigas. In...
Tipo: Text Palavras-chave: Defensin; Antimicrobial peptide; Hemocytes; Pacific oyster; Marine invertebrate; Mollusc bivalve.
Ano: 2007 URL: http://archimer.ifremer.fr/doc/2007/publication-2331.pdf
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