College of Life Sciences, Hebei University, Baoding, Hebei, 071002, China; Laboratory for Marine Biology and Biotechnology, Qingdao National Laboratory for Marine Science and Technology, Qingdao, 266000, China; Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, Qingdao, 266071, China.
College of Life Sciences, Hebei University, Baoding, Hebei, 071002, China.
Fish Shellfish Immunol. 2018 Nov;82:378-385. doi: 10.1016/j.fsi.2018.08.046. Epub 2018 Aug 23.
The gC1qR is a ubiquitously expressed cell protein that interacts with the globular heads of C1q (gC1q) and many other ligands. In this study, one gC1qR homolog gene was obtained from Exopalaemon carinicauda and named EcgC1qR. The complete nucleotide sequence of EcgC1qR contained a 774 bp open reading frame (ORF) encoding EcgC1qR precursor of 257 amino acids. The deduced amino acid sequence of EcgC1qR revealed a 55-amino-acid-long mitochondrial targeting sequence at the N-terminal and a mitochondrial acidic matrix protein of 33 kDa (MAM33) domain. The genomic organization of EcgC1qR gene showed that EcgC1qR gene contained five exons and four introns. EcgC1qR could express in all of the detected tissues and its expression was much higher in hepatopancreas and hemocytes. The expression of EcgC1qR in the hepatopancreas of prawns challenged with Vibrio parahaemolyticus and Aeromonas hydrophila changed in a time-dependent manner. The expression of EcgC1qR in prawns challenged with V. parahaemolyticus was up-regulated at 6 h (p < 0.05), and significantly up-regulated at 12 h and 24 h (p < 0.01), and then returned to the control levels at 48 h post-challenge (p > 0.05). At the same time, the expression in Aeromonas-challenged group was significantly up-regulated at 6, 12 and 24 h. The recombinant EcgC1qR could inhibit the growth of two tested bacteria. In addition, we successfully deleted EcgC1qR gene through CRISPR/Cas9 technology and it was the first time to obtain the mutant of gC1qR homolog gene in crustacean. It's a great progress to study the biological function of gC1qR in crustacean in future.
gC1qR 是一种广泛表达的细胞蛋白，可与 C1q 的球形头部（gC1q）和许多其他配体相互作用。本研究从脊尾白虾（Exopalaemon carinicauda）中获得了一个 gC1qR 同源基因，并将其命名为 EcgC1qR。EcgC1qR 的完整核苷酸序列包含一个 774bp 的开放阅读框（ORF），编码 257 个氨基酸的 EcgC1qR 前体。EcgC1qR 的推导氨基酸序列显示 N 端有一个 55 个氨基酸长的线粒体靶向序列和一个 33kDa 的线粒体酸性基质蛋白（MAM33）结构域。EcgC1qR 基因的基因组组织表明，EcgC1qR 基因包含五个外显子和四个内含子。EcgC1qR 可在所有检测到的组织中表达，在肝胰腺和血细胞中的表达水平更高。在对凡纳滨对虾和哈维弧菌进行攻毒后，EcgC1qR 在对虾肝胰腺中的表达呈时间依赖性变化。凡纳滨对虾感染哈维弧菌后，EcgC1qR 的表达在 6 小时（p<0.05）时上调，在 12 小时和 24 小时（p<0.01）时显著上调，然后在攻毒后 48 小时（p>0.05）时恢复到对照水平。同时，在哈维弧菌攻毒组中，其表达在 6、12 和 24 小时时显著上调。重组 EcgC1qR 可抑制两种受试细菌的生长。此外，我们通过 CRISPR/Cas9 技术成功敲除了 EcgC1qR 基因，这是首次在甲壳动物中获得 gC1qR 同源基因的突变体。这是未来研究甲壳动物 gC1qR 生物学功能的一个重大进展。
