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Mol Immunol ; 59(1): 64-70, 2014 May.
Artigo em Inglês | MEDLINE | ID: mdl-24491489

RESUMO

B cell activating factor (BAFF), a ligand belonging to the tumor necrosis factor (TNF) family is critical to B cell survival, proliferation, maturation and immunoglobulin secretion. In this study, the yellow grouper (Epinephelus awoara) BAFF (designated EaBAFF) gene was cloned using RT-PCR and RACE (rapid amplification of cDNA ends) techniques. The full-length EaBAFF was 1442bp and contained an open reading frame of 780bp encoding a putative protein of 259 amino acids. Amino acids sequence comparison indicated that EaBAFF possessed the TNF signature. The soluble BAFF (EasBAFF) had been cloned into pET28a. SDS-PAGE and Western blotting analysis confirmed that the soluble fusion protein His-EasBAFF was efficiently expressed in Escherichia coli BL21 (DE3). In vitro, the WST-8 assay indicated that EasBAFF was not only able to promote the survival/proliferation of yellow grouper splenic lymphocytes but also able to promote the survival/proliferation of mouse splenic B cells. Our findings may provide valuable information for research into the immune system of E. awoara and EasBAFF may serve as a potential immunologic factor for enhancing immunological efficacy in fish.


Assuntos
Fator Ativador de Células B/genética , Proteínas de Peixes/genética , Expressão Gênica , Perciformes/genética , Sequência de Aminoácidos , Animais , Fator Ativador de Células B/classificação , Fator Ativador de Células B/metabolismo , Linfócitos B/metabolismo , Western Blotting , Clonagem Molecular , Proteínas de Peixes/metabolismo , Camundongos , Microscopia Confocal , Dados de Sequência Molecular , Perciformes/metabolismo , Filogenia , Ligação Proteica , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Homologia de Sequência de Aminoácidos , Baço/citologia , Transcriptoma
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