红鳍东方鲀3个白介素基因的序列特征及表达分析

本试验分析体质量(190.34±2.13)g的红鳍东方鲀白介素基因IL-1b、IL-8和IL-10的序列特征,检测其在红鳍东方鲀脑、鳃、心脏、肌肉、肝脏和脾脏中的表达,以及每尾红鳍东方鲀注射密度1×10^7 cfu/mL的哈维氏弧菌菌液0.1 mL后0、12、24、48 h在肝脏和脾脏中的表达。试验结果表明,IL-1b、IL-8和IL-10基因的序列全长分别为771、822 bp和844 bp。3个基因在健康成鱼脑、鳃、心脏、肌肉、肝脏、脾脏中均有表达,其中肝脏和脾脏表达显著(P<0.05)。感染哈维氏弧菌后12 h,IL-1b基因在肝脏中显著上调,12 h和48 h在脾脏中显著升高(P<0.05)。感染哈维氏弧菌后12 h和24 h IL-8基因在肝脏和脾脏中均显著上调(P<0.05);而感染后24 h和48 h IL-10基因在肝脏中显著升高,在脾脏中12 h和48 h显著升高(P<0.05)。笔者首次探讨红鳍东方鲀IL-1b、IL-8和IL-10基因的序列特性及表达特征,为研究3个基因的原核表达提供了试验信息,将为进一步深入探究重组蛋白活性及其在红鳍东方鲀健康养殖中的应用奠定基础。

Characterization and Expression Profiles of Three Interleukin Genes in Tiger Puffer Takifugu rubripes

The sequence characteristics of the interleukin genes IL-1 b,IL-8 and IL-10 were investigated in tiger puffer Takifugu rubripes with body weight of(190.34±2.13)g,and the expression of interleukin genes was detected in the brain,gills,heart,muscle,liver and spleen of tiger puffer.Changes in expression level of the interleukin genes IL-1 b,IL-8 and IL-10 in liver and spleen were monitored in tiger puffer injected with Vibrio harveyi solution at a dose of 0.1 mL/individual at a concentration of 1×10^7 cells/mL 0 h,12 h,24 h,and 48 h after administration.It was found that the sequence length was 771 bp in IL-1 b,822 bp in IL-8 and 844 bp in IL-10.The three genes were all expressed in heart,brain,gills,muscle,liver,and spleen of normal tiger puffer,significantly higher expression levels in the liver and spleen(P<0.05).In the challenged tiger puffer,IL-1 b gene was shown to be significantly elevated expression level in the liver at 12 h,and in the spleen at 12 h and 48 h post infection with V.harveyi(P<0.05).IL-8 gene expression level was increased significantly in the liver and the spleen at 12 h and 24 h(P<0.05);while IL-10 gene was shown to be significantly expressed in the liver at 24 h,and 48 h,and spleen at 12 h and 48 h(P<0.05).The analysis of sequence characteristics and expression profiles of IL-1 b,IL-8 and IL-10 genes in tiger puffer for the first time provided experimental information for the prokaryotic expression and recombinant proteins activities study of these genes,and the foundation for the application of healthy cultivation of tiger puffer.