虾夷马粪海胆TNFAIP3基因的克隆与表达分析
Cloning and Expression Analysis of TNFAIP3 Gene in Strongylocentrotus intermedius
DOI: 10.12677/AMS.2018.54017, PDF,    国家自然科学基金支持
作者: 刘圣美, 陈亚东, 常亚青, 王秀利, 仇雪梅, 刘 洋*：大连海洋大学，农业部北方海水增养殖重点实验室，辽宁 大连
关键词: 虾夷马粪海胆；SiTNFAIP3；先天免疫；负调控；Strongylocentrotus intermedius； SiTNFAIP3； Innate Immunity； Negative Regulation
摘要: 采用cDNA末端快速扩增(RACE)技术克隆获得一条虾夷马粪海胆(Strongylocentrotus intermedius) SiTNFAIP3基因cDNA全长序列。实时定量PCR检测了SiTNFAIP3基因在组织中的分布及在经脂多糖(LPS)、肽聚糖(PGN)、聚肌胞苷酸(poly I: C)、强壮弧菌(Vibrio fortis)和葡聚糖(WGP)刺激后的表达情况。结果表明：SiTNFAIP3的cDNA全长为3990 bp，5’-UTR为209 bp，开放阅读框为3567 bp，3’-UTR为214 bp，编码1188个氨基酸，具有1个OTU结构域和8个ZnF-A20结构域；qRT-PCR结果显示SiTNFAIP3在检测的各组织中均有表达，卵巢中的表达量显著高于其它组织(p < 0.05)；经PGN、V. fortis刺激后，体腔细胞中SiTNFAIP3表达量显著上升，在刺激后12 h达到最高值(p < 0.05)；经LPS、poly I: C刺激后，体腔细胞中SiTNFAIP3表达变化较小；而经WGP刺激后，体腔细胞中SiTNFAIP3表达出现下调。研究表明，SiTNFAIP3基因参与了虾夷马粪海胆的NLR、TLR和RLR信号通路，但主要参与了NLR信号通路。SiTNFAIP3基因在海胆的先天免疫中发挥重要作用。
Abstract: Using rapid amplification of cDNA ends (RACE) cloned the cDNA full-length sequence of SiTNFAIP3 gene in Strongylocentrotus intermedius. We detected the distribution of SiTNFAIP3 gene in tissues and the expression of SiTNFAIP3 gene stimulated by lipopolysaccharide (LPS), Peptidoglycan (PGN), Polyinosine (poly I: C), Vibrio fortis and glucan (WGP) use real-time quantitative PCR. The results showed that the cDNA full-length of SiTNFAIP3 was 3990 bp, 5’-UTR was 209 bp, open reading frame was 3567 bp, 3’-UTR was 214 bp, encoding 1188 amino acids, with one OTU domain and eight ZnF-A20 domains; qRT-PCR results showed that SiTNFAIP3 was expressed in all detected tissues, and the expression level of SiTNFAIP3 in ovaries was significantly higher than that in other tissues (p < 0.05). After stimulation by PGN and V. fortis, the expression of SiTNFAIP3 increased significantly in coelomic cells, reaching the highest level 12 hours after stimulation (p < 0.05); and after LPS and poly I: C stimulation, the expression of SiTNFAIP3 changed slightly in coelomic cells; but after WGP stimulation, the expression of SiTNFAIP3 gene in coelomic cells decreased. Our studies showed that the SiTNFAIP3 gene was involved in the NLR, TLR and RLR signaling pathway of S. intermedius. It plays an important role in innate immunity of sea urchins.
文章引用：刘圣美, 陈亚东, 常亚青, 王秀利, 仇雪梅, 刘洋. 虾夷马粪海胆TNFAIP3基因的克隆与表达分析[J]. 海洋科学前沿, 2018, 5(4): 144-154. https://doi.org/10.12677/AMS.2018.54017

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