【摘要】 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。更多还原

【Abstract】 The lipopolysaccharide-induced TNF-α factor( LITAF) gene was identified from humphead snapper( Lutjanus sanguineus) spleen using homologous cloning and RACE PCR,designated as Ls-LITAF.The full length c DNA of Ls-LITAF was 815 bp,contained an ORF of 447 bp encoding a polypeptide of 148 amino acids with predicted molecular mass of 15. 7 ku. Sequence alignment showed that Ls-LITAF possessed a typical LITAF domain with a CXXC and( H) x Cxx C motif,shared high similarities with other fish LITAF proteins. Phylogenetic analysis indicated that Ls-LITAF clustered closely with known fish LITAFs. Ls-LITAF was distributed in all analyzed tissues with higher expressions in gill,spleen,skin and head kidney. After Vibrio harveyi infection,the expression levels of Ls-LITAF were up-regulated significantly in spleen and head kidney. These results provided a reference to further investigate the roles of Ls-LITAF in immune response of Lutjanus sanguineus.更多还原