title: Isolation and characterization of LIGHT (TNFSF14) gene homologue in zebrafish (Danio rerio).
abstract: The tumor necrosis factor superfamily (TNFSF) proteins are cytokines involved in many biological processes. In this study, the TNF superfamily member 14 (TNFSF14, LIGHT) has been isolated from zebrafish Danio rerio (designated zLIGHT). The full-length open reading frame (ORF) of zLIGHT cDNA consists of 708 bp encoding a protein of 235 amino acids. The zLIGHT open reading frame (ORF) genomic sequence consists of three introns and four exons, is about 9.9 kb in size. Real-time quantitative PCR (qPCR) analysis suggested that zLIGHT was predominantly expressed in zebrafish spleen. The soluble LIGHT (zsLIGHT) had been cloned into the pSUMO vector, SDS-PAGE and Western blotting analysis confirmed that the recombinant protein SUMO-zsLIGHT was efficiently expressed in Escherichia coli BL21 (DE3). Laser scanning confocal microscopy analysis showed that SUMO-zsLIGHT could bind to its receptors on T cells. LIGHT is involved in many important biological effects, including up-regulating proinflammatory chemokines, cytokines, inducing cell death, apoptosis, and enhancing T cell survival. Zebrafish may conduct as a model animal for further research on LIGHT.
authors.affiliation: Jiangsu Province Key Laboratory for Molecular and Medical Biotechnology, Life Sciences College, Nanjing Normal University, Nanjing 210046, China.
authors.forename: Ai-ying
authors.initials: AY
authors.lastname: Tian
authors.forename: Hui-juan
authors.initials: HJ
authors.lastname: Yang
authors.forename: Shi-cheng
authors.initials: SC
authors.lastname: Zhu
authors.forename: Yu-shun
authors.initials: YS
authors.lastname: Zhang
authors.forename: Zheng-an
authors.initials: ZA
authors.lastname: Jiang
authors.forename: Jin-yun
authors.initials: JY
authors.lastname: Song
authors.forename: Hong-zhen
authors.initials: HZ
authors.lastname: Liu
authors.forename: Shuang-quan
authors.initials: SQ
authors.lastname: Zhang
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D005786:Gene Expression Regulation / Q000502:physiology*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011487:Protein Conformation; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D011994:Recombinant Proteins; D053326:Tumor Necrosis Factor Ligand Superfamily Member 14 / Q000235:genetics / Q000378:metabolism*; D015027:Zebrafish
chemical_list: D012333:RNA, Messenger; D011994:Recombinant Proteins; D053326:Tumor Necrosis Factor Ligand Superfamily Member 14