花鲈肿瘤坏死因子基因cDNA的克隆、分析与表达  

Cloning and expression analysis of tumor necrosis factor α in Japanese sea perch Lateolabrax japonicus

作　　者：邱丽华[1] 宋林生[1] 蔡中华[1] 吴龙涛[1] 江世贵[2] 

机构地区：[1]中国科学院海洋研究所实验海洋生物学开放实验室,山东青岛266071 [2]中国水产科学研究院南海水产研究所,广东广州510300

出　　处：《中国水产科学》2003年第5期370-375,共6页Journal of Fishery Sciences of China

基　　金：国家"八六三"高技术研究发展计划项目(2001AA628180)

摘　　要：采用同源克隆法,结合锚定PCR技术,获得了花鲈(Lateolahraxjaponicus)TNFα基因cDNA的部分序列。BLAST分析表明,该序列与其他鱼类的TNFα基因具有很高的相似性与同源性。同时利用RT PCR技术,对该基因在鱼体内不同组织之间的表达差异进行了分析研究,结果表明,该基因在花鲈免疫器官头肾、脾脏、肝脏中的表达较强,而在脑中的表达较弱,在肌肉中几乎不表达。而且在经特异性病原刺激前、后的组织对比分析中,发现鱼体经LPS(lipopolysaccharide)刺激后,目的基因在免疫器官中的表达明显增强。该基因的克隆与表达为进一步深入研究海水鱼类的抗逆机理以及指导鱼类的遗传选育和遗传改良都具有重要的理论意义。Tumor necrosis factor α (TNFα)is an inflammatory cytokine produced by monocytes or macrophages during acute inflammation and is responsible for a diverse range of signaling events within cells. The partial cDNA of TNFα in Japanese sea perch (Lateolabrax japonicus) was cloned and sequenced with the method of homology cloning. The nucleotide and the deduced protein sequence of the cloned cDNA shows a high degree of homology (>80%) with other known TNFα gene of the fishes, such as Oncorhychus mykiss (89%), Paralichthys olivaceus (95%), Salvelinus fontinalis (87%), Ictalurus punctatus (92%), Sparus aurata (86%), and Pagrus major (84%). The expression analysis using RTPCR shows that the sea perch TNFα gene is constitutively expressed in the headkidney and spleen of the unstimulated fish. The sea perch TNFα gene expression in various tissues can be up-regulated by stimulation with lipoplysaccharide (LPS). The TNFα gene can be expressed in brain after the stimulation with LPS. In the stimulated tissues (headkidney, liver, spleen, brain, and muscle), a strong TNFα expression happens in the headkidney, and follows the spleen and the liver. But there is few expression in the muscle. 

关 键 词：花鲈 肿瘤坏死因子 RT—PCR表达 同源克隆 TNFΑ CDNA 序列 抗逆机理 

分 类 号：Q959.483[生物学—动物学] Q786