cDNA cloning, characterization, and expression analysis of the Rac1 and Rac2 genes from Cynoglossus semilaevis
cDNA cloning, characterization, and expression analysis of the Rac1 and Rac2 genes from Cynoglossus semilaevis
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半滑舌鱼 Rac1 和 Rac2 基因的 cDNA 克隆、表征和表达分析
DOI：
10.1016/j.fsi.2018.11.006
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发表时间：
2019
影响因子：
4.7
通讯作者：
Chen Songlin
中科院分区：
文献类型：
--
作者：
Xiu Yunji;Zhang Hongxiang;Wang Shuangyan;Gan Tian;Wei Min;Zhou Shun;Chen Songlin
Rac1 和 Rac2 属于 Rho GTPase 小家族，在免疫反应中发挥着重要作用。在本研究中，从半滑舌鱼中克隆了aRac1同源物（CsRac1）和aRac2同源物（CsRac2）。 CsRac1和CsRac2cDNA全长分别为1219 bp和1047 bp。 CsRac1和CsRac2均含有579 bp的开放阅读框(ORF)，编码192个氨基酸的推定蛋白。 CsRac1和CsRac2的预测分子量分别为21.41kDa和21.35kDa，理论等电点分别为8.50和7.91。序列分析表明，CsRac1和CsRac2的氨基酸均检测到保守的RHO结构域。同源分析表明CsRac1和CsRac2与不同物种的其他对应物具有高度保守性。 CsRac1和CsRac2转录本在组织中分布广泛，其中CsRac1和CsRac2分别在肝脏和鳃中表达量最高。哈维弧菌攻击后不同时间点肝脏和鳃组织中CsRac1和CsRac2的表达水平出现波动。具体而言，CsRac1和CsRac2在注射后48小时和96小时显着上调。此外，敲除细胞系（TSHKC）中的CsRac1和CsRac2可降低CsPAK1、CsIL1-β和CsTNF-α的表达。目前的数据表明CsRac1和CsRac2可能在半滑舌鳎的天然免疫中发挥重要作用。
Rac1 and Rac2, belonging to the small Rho GTPase family, play an important role during the immune responses. In this study, aRac1homolog (CsRac1) and aRac2homolog (CsRac2) were cloned from theCynoglossus semilaevis. The full-length ofCsRac1andCsRac2cDNA was 1219 bp and 1047 bp, respectively. BothCsRac1andCsRac2contain a 579 bp open reading frame (ORF) which encoding a 192 amino acids putative protein. The predicted molecular weight ofCsRac1andCsRac2was 21.41 kDa and 21.35 kDa, and their theoretical pI was 8.50 and 7.91, respectively. Sequence analysis showed that the conserved RHO domain was detected both from amino acid ofCsRac1andCsRac2. Homologous analysis showed thatCsRac1andCsRac2share high conservation with other counterparts from different species. TheCsRac1andCsRac2transcript showed wide tissue distribution, in whichCsRac1andCsRac2exhibit the highest expression level in liver and gill, respectively. The expression level ofCsRac1andCsRac2fluctuated in the liver and gill tissues at different time points after challenged byVibrio harveyi. Specifically,CsRac1andCsRac2were significantly up-regulated at 48 h and 96 h post injection. Moreover, the knocking down ofCsRac1andCsRac2in cell line (TSHKC) reduced the expression ofCsPAK1, CsIL1-βandCsTNF-α. The present data suggests thatCsRac1andCsRac2might play important roles in the innate immunity of half-smooth tongue sole.