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Min, C, Liang, Z, Cui, X, Wang, Q, Chen, Y and Zhang, S (2012) Molecular cloning and partial functional characterization of a proliferation inducing ligand (APRIL) in zebrafish (Danio rerio). Dev. Comp. Immunol. 37:202-6

Here we describe the identification of a Danio rerio homologue of a proliferation-inducing ligand (APRIL) of the TNF family (designated zAPRIL). Sequence analysis showed that the open reading frame of zAPRIL consists of 600 bases encoding a protein of 199-amino acids. Recombinant soluble APRIL (zsAPRIL) was constructed consisting of fluorescence-enhanced green fluorescent protein (EGFP) and cloned into a pET28a vector. SDS-PAGE and western blotting analysis indicated a high-level expression of soluble EGFP/zsAPRIL protein in Escherichia coli BL21 (DE3). Observation by confocal microscopy demonstrated that EGFP/zsAPRIL could successfully bind to the surface receptors of zebrafish lymphocytes. In vitro survival analysis revealed that purified EGFP/zsAPRIL was able to promote the survival of zebrafish lymphocytes in a dose-dependent manner. The biological role of APRIL does not seem to be restricted to proliferation induction. Zebrafish may could served as a model organism for further study of APRIL.

PubMed Online version:10.1016/j.dci.2011.12.002

Animals; Cell Proliferation; Cell Survival; Cells, Cultured; Cloning, Molecular; Lymphocytes/physiology; Molecular Sequence Data; Phylogeny; Recombinant Fusion Proteins/genetics; Recombinant Fusion Proteins/metabolism; Recombinant Fusion Proteins/physiology; Sequence Analysis, DNA; Sequence Homology, Amino Acid; Spleen/cytology; Tumor Necrosis Factor Ligand Superfamily Member 13/genetics; Tumor Necrosis Factor Ligand Superfamily Member 13/metabolism; Tumor Necrosis Factor Ligand Superfamily Member 13/physiology; Zebrafish; Zebrafish Proteins/genetics; Zebrafish Proteins/metabolism; Zebrafish Proteins/physiology