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尼罗罗非鱼TRIM16和TRIM25基因的克隆及表达分析
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摘要
泛素连接酶(E3)家族在脊椎动物的先天免疫中发挥了重要作用。为了研究TRIM16(tripartitemotif containing 16)和TRIM25(tripartite motif containing 25)在尼罗罗非鱼(Oreochromisniloticus)先天免疫中的作用,本实验通过RT-PCR和末端快速扩增(RACE)方法获得尼罗罗非鱼TRIM16和TRIM25的cDNA全序列,并分析其基因结构和组织表达特征。结果表明:TRIM16的cDNA全长为2314bp,ORF长1677bp可编码588个氨基酸,TRIM25的cDNA全长为2748bp,ORF长1677bp可编码588个氨基酸,二者基因组序列中均无内含子。推测氨基酸序列结构,TRIM16和TRIM25均具有TRIM家族的RING finger结构域、B-box结构域和卷曲螺旋结构域等保守结构域。组织分析表明:在本实验所检测的11个组织和器官中,TRIM16和TRIM25均在血液中表达量最高。人工感染无乳链球菌(Streptococcus agalactiae)后,TRIM16和TRIM25在各检测组织中的表达量都有明显上调。
The ubiquitin ligase(E3) family plays an important role in the innate immunity of vertebrates.In the present study, we evaluated the immune function of TRIM16 and TRIM25 in Nile tilapia.The full-length cDNA sequences of TRIM16 and TRIM25 from Nile tilapia were obtained by RT-PCR and rapid amplification of cDNA ends polymerase chain reaction(RACE-PCR). The gene structure and the tissue expression of TRIM16 and TRIM25 were also studied. Results showed that the TRIM16 gene was 2314-bp in length with an open reading frame(ORF) of 1677-bp encoding 588 amino acid residues and with no intron. TRIM25 gene was2748-bp in length with an open reading frame(ORF) of 1677-bp encoding 588 amino acid residues and with no intron.Gene structureindicated that the TRIM16 and TRIM25 contained the conserved structure of TRIM family.Tissue distribution analysis revealed the presence of TRIM16 and TRIM25 transcript in eleven normal tissues. The TRIM16 and TRIM25 presented the highest expression level in blood. Upon stimulation with intraperitoneal injection of Streptococcus agalactiae,the expression levels of TRIM16 and TRIM25 were up-regulated in all tissue.
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