http://http://dspace.unitus.it:80 The DSpace digital repository system captures, stores, indexes, preserves, and distributes digital research material. Fri, 24 May 2013 07:41:21 GMT 2013-05-24T07:41:21Z http://hdl.handle.net/2067/1688 Title: Molecular and structural characterisation of a macrophage migration inhibitory factor from sea bass (Dicentrarchus labrax L.) Authors: Buonocore, Francesco; Randelli, Elisa; Facchiano, Angelo M.; Pallavicini, Alberto; Modonut, Martina; Scapigliati, Giuseppe Abstract: The macrophage migration inhibitory factor (MIF) is a cytokine produced in numerous cell types, mainly T lymphocytes and macrophages, in response to inflammatory stimuli. In this paper we report the identification of a cDNA encoding a MIF molecule from sea bass (Dicentrarchus labrax L.), its expression analysis and its 3D structure obtained by template-based modelling. The sea bass MIF cDNA consists of 609 bp that translates in one reading frame to give the entire molecule containing 115 amino acids. The sequence contains three cysteine residues in conserved positions compared to human MIF and most Teleost fishes, with the exception of zebrafish and carp. The Cys57-Ala58-Leu59-Cys60 motif, present inside the stretch important for JAB1-interaction and mediator of the thiol-protein oxidoreductase activity of MIF, is conserved in sea bass, together with the Pro2 residue that is crucial for the tautomerase catalytic activity. Real time PCR analyses revealed that MIF is constitutively expressed in all selected tissues and organs, with the highest mRNA level observed in thymus. MIF expression was induced after 4 hours in vitro stimulation of head kidney leukocytes with LPS and decreased after 24 hours. The predicted 3D model of sea bass MIF has been used to verify the presence of structural requirements for its known biological activities. Description: L'articolo è disponibile sul sito dell'editore http://www.sciencedirect.com/ Thu, 31 Dec 2009 23:00:00 GMT http://hdl.handle.net/2067/1688 2009-12-31T23:00:00Z http://hdl.handle.net/2067/1690 Title: Searching for immunomodulatory sequences in sea bass (Dicentrarchus labrax L.): Transcripts analysis from thymus Authors: Pallavicini, Alberto; Randelli, Elisa; Modonut, Martina; Casani, Daniela; Scapigliati, Giuseppe; Buonocore, Francesco Abstract: The thymus is a key organ of the immune system in most vertebrates and, for this reason, it has been used in this paper for the generation of a normalized cDNA library from sea bass (Dicentrarchus labrax), one of the most extensively cultured species in South Mediterranean aquaculture. A total of 1,632 ESTs from this library were initially analyzed for sequence quality and vector sequences and, after this control, 1,264 (77% of total clones sequenced) high-quality ESTs were further processed. The total collection of Dicentrarchus labrax thymus ESTs has been deposited in the EBI-GenBank-DBJ database (GenBank accession numbers from FN565576 to FN566839). The functional classification of ESTs was performed by Gene Ontology and KEGG annotation and, successively, the sequences were analysed using the ImmunomeBase software to identify potentially immuno related genes. Using this approach, we found about 100 putative genes involved in immune system responses, most new in sea bass, that were analysed most in detail. Some of the potentially interesting genes identified by these in silico analyses were studied by real-time PCR to verify their expression both at basal level and after in vitro stimulation of sea bass head kidney leukocytes. The used strategy has been confirmed as a good approach to discovery new immuno-related genes and to improve the knowledge of specific markers that could help the discrimination of T cell subpopulations in sea bass and, in general, in Teleosts. Description: L'articolo è disponibile sul sito dell'editore http://www.sciencedirect.com/ Thu, 31 Dec 2009 23:00:00 GMT http://hdl.handle.net/2067/1690 2009-12-31T23:00:00Z