Molecular cloning, expression and functional analysis of B-cell activating factor (BAFF) in yellow grouper, Epinephelus awoara
- Authors
- Xiao, W., Long, W., Liu, G.Y., Sui, C.L., Guo, X.R., Tian, A., Ji, C.B., Cui, X.W., and Zhang, S.Q.
- ID
- ZDB-PUB-140728-40
- Date
- 2014
- Source
- Molecular immunology 59(1): 64-70 (Journal)
- Registered Authors
- Keywords
- none
- MeSH Terms
-
- Amino Acid Sequence
- Animals
- B-Cell Activating Factor/classification
- B-Cell Activating Factor/genetics*
- B-Cell Activating Factor/metabolism
- B-Lymphocytes/metabolism
- Blotting, Western
- Cloning, Molecular
- Fish Proteins/genetics*
- Fish Proteins/metabolism
- Gene Expression*
- Mice
- Microscopy, Confocal
- Molecular Sequence Data
- Perciformes/genetics*
- Perciformes/metabolism
- Phylogeny
- Protein Binding
- Reverse Transcriptase Polymerase Chain Reaction
- Sequence Homology, Amino Acid
- Spleen/cytology
- Transcriptome
- PubMed
- 24491489 Full text @ Mol. Immunol.
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 1442 bp and contained an open reading frame of 780 bp 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.