Molecular structure, expression analysis and functional characterization of APRIL (TNFSF13) gene in bat (Vespertilio superans Thomas)
- 作者：Fengtao You ; Lidan Zhou ; Xuanxuan Liu ; Jie Fan ; Zhen Ke ; Wenhua Ren ; youfengtao1014@yahoo.cn
- 关键词：APRIL ; A proliferation-inducing ligand ; bp ; base pair(s) ; BAFF ; B cell activating factor ; BCMA ; B cell maturation antigen ; cDNA ; DNA complementary ; DMSO ; dimethylsulfoxide ; dNTP ; deoxyribonucleoside triphosphate ; HSPG ; heparin sulfate proteoglycan ; IPTG
- 刊名：Gene
- 出版年：2012
- 期刊代码：73_03781119
- 类别：bio
- 出版时间：1 May, 2012
- 卷：498
- 期：2
- 页码：196-202
- 文件大小：1058 K
摘要
A proliferation-inducing ligand (APRIL) is a novel member of the tumor necrosis factor (TNF) superfamily, which is involved in immune regulation. In the present study, the full-length cDNA of APRIL (designated bAPRIL) from bat was cloned using RT-PCR and its biological activities have been characterized. The open reading frame (ORF) of this cDNA consists of 753 bases, encoding a protein of 250 amino acids. This protein was found to contain a predicted transmembrane domain, a putative furin protease cleavage site, and a typical TNF homology domain corresponding to other, known APRIL homologs. Real-time quantitative PCR (qPCR) analysis indicated that bAPRIL mRNA was predominantly expressed in bat lymphoid tissue spleen. The SUMO-bsAPRIL was efficiently expressed in Escherichia coli BL21 (DE3) and confirmed by SDS-PAGE and Western blot analysis. Laser scanning confocal microscopy analysis showed that bsAPRIL could bind to its receptors on B cells. In vitro, MTT assays indicated that bsAPRIL could promote the survival/proliferation of mouse splenic B cells grown with anti-mouse IgM. These findings indicate that bsAPRIL plays an important role in the survival and proliferation of B cells and has functional cross-reactivity among mammalians. The present findings may provide valuable information for research into the immune system of the bat.