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孙梦洁, 陈亚东, 高杰, 江炎亮, 张雪, 佘定懿, 许式见, 胡谋, 赵爱云, 沙珍霞. 俄罗斯鲟补体C7基因的克隆及表达分析[J]. 水生生物学报, 2019, 43(4): 763-770. DOI: 10.7541/2019.090
引用本文: 孙梦洁, 陈亚东, 高杰, 江炎亮, 张雪, 佘定懿, 许式见, 胡谋, 赵爱云, 沙珍霞. 俄罗斯鲟补体C7基因的克隆及表达分析[J]. 水生生物学报, 2019, 43(4): 763-770. DOI: 10.7541/2019.090
SUN Meng-Jie, CHEN Ya-Dong, GAO Jie, JIANG Yan-Liang, ZHANG Xue, SHE Ding-Yi, XU Shi-Jian, HU Mou, ZHAO Ai-Yun, SHA Zhen-Xia. CLONING AND EXPRESSION ANALYSIS OF THE COMPLEMENT COMPONENT C7 IN RUSSIAN STURGEON (ACIPENSER GUELDENSTAEDTI)[J]. ACTA HYDROBIOLOGICA SINICA, 2019, 43(4): 763-770. DOI: 10.7541/2019.090
Citation: SUN Meng-Jie, CHEN Ya-Dong, GAO Jie, JIANG Yan-Liang, ZHANG Xue, SHE Ding-Yi, XU Shi-Jian, HU Mou, ZHAO Ai-Yun, SHA Zhen-Xia. CLONING AND EXPRESSION ANALYSIS OF THE COMPLEMENT COMPONENT C7 IN RUSSIAN STURGEON (ACIPENSER GUELDENSTAEDTI)[J]. ACTA HYDROBIOLOGICA SINICA, 2019, 43(4): 763-770. DOI: 10.7541/2019.090

俄罗斯鲟补体C7基因的克隆及表达分析

CLONING AND EXPRESSION ANALYSIS OF THE COMPLEMENT COMPONENT C7 IN RUSSIAN STURGEON (ACIPENSER GUELDENSTAEDTI)

  • 摘要: 为研究俄罗斯鲟(Acipenser gueldenstaedti)补体C7基因(AgC7)的功能, 采用RACE (Rapid-amplification of cDNA ends)技术, 获得AgC7的cDNA全长序列为3103 bp, 包括开放阅读框(Open Read Frame, ORF) 2502 bp, 编码833个氨基酸, 5′-UTR (5′ untranslated region)和3′-UTR的长度分别为44和554 bp。同源性分析表明, AgC7与其他鱼类补体C7如斑点雀鳝(Lepisosteus oculatus)、斑点叉尾鲴(Ictalurus punctatus)、斑马鱼(Danio rerio)、大西洋鲑(Salmo salar)、尼罗罗非鱼(Oreochromis niloticus)氨基酸序列的一致性分别为56%、46%、49%、49%、47%, 且都具有TSP1、LDLa、FIMAC和MACPF保守结构域。荧光定量qRT-PCR (quantitative Real-time PCR)结果表明, AgC7在血液、脑、鳃、性腺、心脏、头肾、肠、肝、肌肉、皮肤、脾、胃和后肾组织中均可表达, 且在肠中表达水平最高; 用含有壳寡糖的饲料饲喂俄罗斯鲟60d后, AgC7在这13种组织中表达均有增加, 在肠中增加量最高, 约为对照组的1.51倍; 嗜水气单胞菌(Aeromonas hydrophila)可诱导AgC7, 在血液、鳃、头肾、肠、肝和脾6种组织中瞬时上调表达, 随着病原菌感染时间增加, 基因表达量逐渐降低恢复至正常水平, 其中, 鳃中基因表达量的上调趋势最明显, 最大表达量出现在感染后6h, 为对照组的41.30倍, 12h后基因表达下降并恢复至正常水平, 表明AgC7基因可能参与了俄罗斯鲟抗细菌感染的免疫应答。俄罗斯鲟AgC7具有典型的补体C7基因特征, 且壳寡糖刺激和病原感染后均可引起AgC7基因表达变化, 补体C7可能参与了俄罗斯鲟的免疫应答。

     

    Abstract: This study cloned the full-length of complement C7 cDNA by RACE (rapid-amplification of cDNA ends) from Russian sturgeon (Acipenser gueldenstaedti). It contained 3103 bp, including a 2502 bp open reading frame (ORF) encoding 833 dedueced amino acids, a 44 bp 5′-UTR and a 554 bp 3′-UTR. Multiple sequence alignment revealed that AgC7 proteins were well conserved with the typical structural motifs such as TSP1, LDLa, FIMAC, MACPF and had the sequence identity with Lepisosteus oculatus (56%), Ictalurus punctatus (46%), Danio rerio (49%), Salmo salar (49%) and Oreochromis niloticus (47%), respectively. AgC7 gene is constitutively expressed in all evaluated tissues with the highest expression in intestine and the weakest level in muscle. Chitosan oligosaccharide induced the expression of AgC7 gene to a certain degrees in all tissues with the highest increase in intestine (1.51 times). After infection with Aeromonas hydrophila, AgC7 gene exhibited distinct time-dependent response patterns in tested tissues with most obvious induction in gills and the maximum expression at 6h after infection (41.30 times). The expression of AgC7 gene decreased rapidly and returned to normal level after 12h. These results suggest that C7 of Russian sturgeon might play an important role in the immune defenses against bacterial infection.

     

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