广东海洋大学学报2019，Vol.39Issue(6)：1-8,8.DOI:10.3969/j.issn.1673-9159.2019.06.001
尼罗罗非鱼淋巴毒素α基因克隆与表达分析
Cloning and Expression Analysis of Gene Lymphotoxin α in Nile Tilapia (Oreochromis Niloticus)
摘要
Abstract
[Objective] To clone and analyze the lymphotoxin alpha gene (LTα), analyze the transcript distribution in different tissues, and to explore the important role of lymphotoxin alpha (LTα) in the antibacterial and antiviral immunity of Oreochromis niloticus. [Method]The full-length cDNA of lymphotoxin a from Nile Tilapia (On-LTα ) was obtained by RACE. The sequence of On-LTα was analyzed by bioinformatics method. The tissue distribution and expression profiles of On-LTα following Streptococcus lactobacillus immunization were investigated by Real-time quantitative PCR. Furthermore, On-LTα transcript levels in the isolated non-specific toxic cells (NCC) after lipopolysaccharide (LPS) and Poly I:C stimulation were also examined.[Results ]On-LTα gene sequence is 2699 bp including a 705 bp open reading frame (ORF) which encodes a polypeptide of 234 amino acids. (GenBank accession number: MK770358). On-LTα protein belonged to the transmembrane protein and possessed a conserved domain of the tumor necrosis factor (TNF) family. Quantification PCR found that On-LTα was constitutively expressed in all 11 tissues tested. In the healthy Nile tilapia, the highest expression being detected in the spleen. The relative expression levels of On-LTα in the spleen were significantly increased during S. agalactia infection. The isolated NCCs cultured in vitro could also induce the significant upregulation of On-LTα under LPS and Poly I: C stimulation. [Conclusion]On-LTαmight be involved in the antimicrobial and antiviral immune responses of tilapia.关键词
尼罗罗非鱼/淋巴毒素α基因/基因克隆/基因表达/非特异性细胞毒性细胞(NCC)Key words
Oreochromis niloticus/gene lymphotoxin alpha/gene clone/gene expression/Non-specific cytotoxic cell (NCC)分类
生物科学引用本文复制引用
邹子鸿,郑琦,黄瑜,汤菊芬,王蓓,鲁义善,简纪常,蔡佳..尼罗罗非鱼淋巴毒素α基因克隆与表达分析[J].广东海洋大学学报,2019,39(6):1-8,8.基金项目
国家自然科学基金(31302226, 31572651) （31302226, 31572651）
广东海洋大学优秀硕士学位论文培育项目(201833) （201833）
