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通过同源克隆和RACE PCR技术从红笛鲷脾脏中鉴定得到了脂多糖诱导的肿瘤坏死因子(LITAF)基因,命名为Ls-LITAF。该基因c DNA全长815 bp,开放阅读框为447 bp,编码148个氨基酸,理论分子量为15.7 ku。序列分析显示Ls-LITAF蛋白具有保守的LITAF结构域,其中包含一个CXXC基序与一个(H)x Cxx C基序。Ls-LITAF蛋白与其他鱼类LITAF蛋白相似度较高,在系统进化树中也与其他鱼类该蛋白聚为一支。Ls-LITAF基因在健康鱼体多种组织均有表达,其中鳃、脾脏、皮肤与头肾的表达量较高;且哈氏弧菌刺激鱼体后,Ls-LITAF在脾脏和头肾中的表达量显著上调。以上研究结果为进一步了解Ls-LITAF在红笛鲷免疫反应中的作用提供了参考。
Lipopolysaccharide-induced tumor necrosis factor (LITAF) gene was identified from the spleen of Red snapper by homologous cloning and RACE PCR and named as Ls-LITAF. The full-length cDNA of this gene is 815 bp in length and contains an open reading frame of 447 bp encoding a protein of 148 amino acids with a theoretical molecular weight of 15.7 ku. Sequence analysis revealed that the Ls-LITAF protein has a conserved LITAF domain that contains one CXXC motif and one (H) x Cxx C motif. Ls-LITAF protein is highly similar to other fish LITAF proteins and clustered with other fish in the phylogenetic tree. Ls-LITAF gene was expressed in many tissues of healthy fish, among which, gill, spleen, skin and head kidney were higher than others; Ls-LITAF gene was expressed in spleen and head kidney after Vibrio harveyi stimulated fish The amount of expression was significantly up-regulated. The above results provide a reference for further understanding the role of Ls-LITAF in the immune response of red snapper.