虾夷马粪海胆LRCH2-Like基因的克隆与表达特征分析
Cloning and Expression Analysis of LRCH2-Like Gene in Strongylocentrotus intermedius
DOI: 10.12677/AMS.2018.52003, PDF,    国家自然科学基金支持
作者: 邹惠冬, 白 莉, 刘圣美, 陈亚东, 周淑红, 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；LRCH2-Like；RACE；实时定量PCR；Strongylocentrotus intermedius； LRCH2-Like； RACE； qRT-PCR
摘要: 本实验采用cDNA末端快速扩增(RACE)技术克隆获得了一条虾夷马粪海胆(Strongylocentrotus intermedius)LRCH2-like基因的全长cDNA序列。SiLRCH2-like基因全长为2446 bp，其中编码区为1644 bp，3’-UTR为609 bp，5’-UTR为193 bp，编码548个氨基酸，预测的蛋白分子量为59.9 KDa，理论等电点为4.07，为酸性蛋白，有1个Na-Ca-ex-superfamily结构域和7个LRR结构域。我们采用实时定量PCR技术分析了SiLRCH2-like基因在各组织中的表达情况。他在所有检测的组织中均有表达，在雌性性腺中表达量最高，其次是肠道、管足，体腔细胞和围口膜中表达量较少，在雄性性腺中几乎检测不到表达。我们也检测了该基因在强壮弧菌(Vibrio fortis)、脂多糖(LPS)、肽聚糖(PGN)、葡聚糖(WGP)、聚肌胞苷酸(polyI:C)五种病原刺激后的体腔细胞中的表达情况。SiLRCH2-like基因在PGN处理后在12 h出现了表达高峰，在WGP处理后出现了表达下调，而其他病原处理后表达上调不明显。研究结果表明，SiLRCH2-like基因参与了虾夷马粪海胆的NLR信号通路，它在海胆的天然免疫系统中发挥了重要的作用。
Abstract: One cDNA of LRCH2-like named SiLRCH2-like was cloned from sea urchin Strongylocentrotus in-termedius through Smart RACE technology. The full-length cDNA sequence of SiLRCH2-like is 2446 bp, with 1644 bp in coding region, 609 bp in 3’-UTR, 193 bp in 5’-UTR, encoding 548 amino acids. Predicted protein molecular weight is 59.9 KDa, and its theoretical isolectric point is 4.07. SiLRCH2-like is an acidic protein that contains a Na-Ca-ex-superfamily domain and 7 leucine-rich repeats (LRRs). SiLRCH2-like was assessed in all tested tissues using quantitative real-time PCR (qRT-PCR). The expression level in female gonad was significantly higher than the others, secondly in gut with tube feet, and lower in coelomocytes with peristomial membrane, but the expression is few in male gonad. Expression distribution in coelomocytes after challenge with Vibrio fortis, LPS, PGN, WGP and polyI:C were also detected qRT-PCR. The expression level of SiLRCH2-like in coelomocytes was strongly up-regulated after challenge with PGN, reaching the highest point at 12 h; the expression level of SiLRCH2-like was weakly down-regulated after challenge with WGP, and showed no significant difference after challenge with others. The findings showed that SiLRCH2- like gene participated in the NOD-like receptor signaling pathway and played an important role in innate immune system of sea urchin.
文章引用：邹惠冬, 白莉, 刘圣美, 陈亚东, 周淑红, 仇雪梅, 刘洋. 虾夷马粪海胆LRCH2-Like基因的克隆与表达特征分析[J]. 海洋科学前沿, 2018, 5(2): 18-28. https://doi.org/10.12677/AMS.2018.52003

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