Jiangsu Province Key Laboratory for Molecular and Medical Biotechnology, Life Sciences College, Nanjing Normal University, Nanjing 210046, China.
Int Immunopharmacol. 2012 Dec;14(4):629-34. doi: 10.1016/j.intimp.2012.09.015. Epub 2012 Oct 3.
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.
肿瘤坏死因子超家族（TNFSF）蛋白是参与多种生物学过程的细胞因子。在本研究中，从斑马鱼（Danio rerio）中分离到了肿瘤坏死因子超家族成员 14（TNFSF14，LIGHT）（命名为 zLIGHT）。zLIGHT cDNA 的全长开放阅读框（ORF）由 708bp 组成，编码 235 个氨基酸的蛋白质。zLIGHT 开放阅读框（ORF）基因组序列由三个内含子和四个外显子组成，大小约为 9.9kb。实时定量 PCR（qPCR）分析表明，zLIGHT 在斑马鱼脾脏中表达丰度较高。已将可溶性 LIGHT（zsLIGHT）克隆到 pSUMO 载体中，SDS-PAGE 和 Western blotting 分析证实重组蛋白 SUMO-zsLIGHT 在大肠杆菌 BL21（DE3）中高效表达。激光扫描共聚焦显微镜分析表明，SUMO-zsLIGHT 可以与 T 细胞上的受体结合。LIGHT 参与许多重要的生物学效应，包括上调促炎趋化因子、细胞因子、诱导细胞死亡、凋亡和增强 T 细胞存活。斑马鱼可能作为 LIGHT 进一步研究的模型动物。
