title: Molecular cloning, expression, bioinformatics analysis, and bioactivity of TNFSF13 (APRIL) in the South African clawed frog (Xenopus laevi): a new model to study immunological diseases.
abstract: TNFSF13 is one of the tumor necrosis factor (TNF) superfamily members that plays important roles in immune homeostasis and proliferation or apoptosis of certain tumor cell lines. This report describes the development of Xenopus laevis TNFSF13 as a model to study its important role in relation to immunological diseases. In brief, TNFSF13 from Xenopus laevis (designated XlTNFSF13) was first amplified by RT-PCR and rapid amplification of cDNA end (RACE) techniques. Bioinformatics analyses revealed the gene structure, three-dimensional structure, and evolutionary relationships. Real-time quantitative PCR (QPCR) analysis identified the tissue distribution of XlTNFSF13 in the major visceral organs. The recombinant plasmid SUMO-XsTNFSF13 was expressed in E. coli Rosseta (DE3). Subsequently, the recombinant protein purified through Ni-NTA affinity chromatography was analyzed by SDS-PAGE and confirmed by Western blot analysis. Laser scanning confocal microscopy analysis revealed the binding activity of pSUMO-XsTNFSF13 to the surface of B cells. WST-8 assays further indicated that purified XsTNFSF13 could cause the survival/proliferation of B cells. In conclusion, we underscore that as a model organism for human disease, Xenopus laevis has been widely used in molecular biology research. Yet while TNFSF13 research in mammalian, fish (e.g., zebrafish), mouse, and human is widely available, studies in the amphibian species are limited. The latter area of OMICS and integrative biology scholarship is directly informed with the present study, with a view to implications for the future study of human immunological diseases.
authors.affiliation: Molecular and Medical Biotechnology and Aquatic Crustacean Diseases, Jiangsu Province Key Laboratory for Life Sciences College, Nanjing Normal University, Nanjing, China.
authors.forename: Xia
authors.initials: X
authors.lastname: Liu
authors.forename: Xue-mei
authors.initials: XM
authors.lastname: Ji
authors.forename: Xi-ning
authors.initials: XN
authors.lastname: Du
authors.forename: Xi-cui
authors.initials: XC
authors.lastname: Zong
authors.forename: Ding-fang
authors.initials: DF
authors.lastname: Liang
authors.forename: Li
authors.initials: L
authors.lastname: Ma
authors.forename: Hai-tao
authors.initials: HT
authors.lastname: Wu
authors.forename: Shuang-quan
authors.initials: SQ
authors.lastname: Zhang
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D053264:B-Cell Activating Factor / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology*; D001402:B-Lymphocytes / Q000276:immunology; D049109:Cell Proliferation; D002470:Cell Survival / Q000235:genetics / Q000276:immunology; D003001:Cloning, Molecular / Q000379:methods; D019295:Computational Biology / Q000379:methods; D018076:DNA, Complementary / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D007154:Immune System Diseases / Q000235:genetics* / Q000276:immunology; D051379:Mice; D008969:Molecular Sequence Data; D010802:Phylogeny; D011485:Protein Binding; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D016415:Sequence Alignment; D014018:Tissue Distribution; D029867:Xenopus Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology*; D014982:Xenopus laevis
chemical_list: D053264:B-Cell Activating Factor; D018076:DNA, Complementary; D011994:Recombinant Proteins; C583528:TNFSF13B protein, Xenopus; D029867:Xenopus Proteins