论文部分内容阅读
为了研究鳜IRAK4生物学特性及其在抗病毒免疫应答中的作用,根据鳜转录组数据中筛选出的IRAK4 unigene序列设计引物,利用SMART-RACE技术克隆得到CDS全长为1389 bp的c DNA(命名为Sc IRAK),编码462个氨基酸,含有1个N端死亡结构域和1个保守的中央蛋白激酶结构域。采用荧光定量RT-PCR方法分析了Sc IRAK4在健康鳜各组织中的表达差异及病毒感染后在脾脏中的表达变化,结果显示,健康鳜中Sc IRAK4在肝脏中表达量最大,与其他组织差异显著,而在血液、脑和胃中表达量最低;传染性脾肾坏死病毒(infectious spleen and kidney necrosis virus,ISKNV)感染鳜后Sc IRAK4的表达量呈现下调趋势,24 h脾脏中的表达量达到最低,为对照组的45%;而鳜弹状病毒(siniperca chuatsi rhabdovirus,SCRV)感染鳜后Sc IRAK4的表达量呈现上调趋势,12 h脾脏中Sc IRAK4的表达量达到最高,为对照组的8.17倍,表明Sc IRAK4在抗ISKNV和SCRV的免疫应答中可能发挥不同的作用。本研究为进一步揭示Sc IRAK4的抗病毒免疫反应机制提供了依据。
In order to study the biological characteristics of IRAK4 and its role in antiviral immune response, primers were designed based on the IRAK4 unigene sequences screened from the transcriptome data. The cDNA of CDS with a total length of 1389 bp was obtained by SMART-RACE Designated as Sc IRAK), encodes 462 amino acids and contains 1 N-terminal death domain and 1 conserved central protein kinase domain. The expression of Sc IRAK4 in healthy tissues was analyzed by fluorescence quantitative RT-PCR. The expression of Sc-IRAK4 in the spleen was also observed. The results showed that the expression of Sc-IRAK4 was the highest in the liver, While the expression level of Sc IRAK4 in infectious spleen and kidney necrosis virus (ISKNV) showed a downward trend. The expression of Sc IRAK4 in 24 h spleen reached the peak The lowest was 45% of the control group. However, the expression of Sc IRAK4 showed an upward trend after infection with siniperca chuatsi rhabdovirus (SCRV), and the expression of Sc IRAK4 reached the peak at 12 h Fold, indicating that Sc IRAK4 may play a different role in the immune response to ISKNV and SCRV. This study provides the basis for further revealing the anti-viral immune response mechanism of Sc IRAK4.