Molecular cloning and partial functional characterization of a proliferation inducing ligand (APRIL) in zebrafish (Danio rerio)

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• 作者：Cui Min^a ; ¹ ; Zhenning Liang^a ; ¹ ; Xianwei Cui^a ; Qi Wang^a ; Yuqing Chen^a ; ^b ; ^c ; ^{yuqingchen515@yahoo.com} ; Shuangquan Zhang^a ; ^b ; ^c ; ^{shuangquanz@yahoo.com}
• 关键词：Zebrafish (Danio rerio) ; APRIL ; cDNA cloning ; Heterologous expression ; Survival assay
• 刊名：Developmental and Comparative Immunology
• 出版年：2012
• 期刊代码：35_0145305x
• 类别：abs
• 出版时间：May, 2012
• 卷：37
• 期：1
• 页码：202-206
• 文件大小：404 K

摘要

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.