| 摘要: |
| 本研究利用本实验室构建的脊尾白虾(Exopalaemon carinicauda)血细胞cDNA文库中的C-型凝集素基因EST序列, 采用cDNA末端快速扩增(rapid amplification of cDNA end, RACE)技术克隆获得脊尾白虾C-型凝集素基因cDNA全长, 命名为EcCTL。该基因全长1285 bp, 包含1041 bp的开放阅读框,编码346个氨基酸组成的蛋白质, 分子量为38.56 kDa, 为甘露糖型凝集素。同源性分析表明, 脊尾白虾EcCTL基因氨基酸序列与秀丽白虾(Palaemon modestus)的同源性最高, 达到91％。荧光定量PCR分析结果表明, EcCTL基因在血细胞、肝胰腺、肌肉等组织中均有表达, 其中在肝胰腺当中的相对表达量最高。感染鳗弧菌(Vibrio anguillarum)和白斑综合症病毒(white spot syndrome virus, WSSV)后6～12 h,脊尾白虾血细胞和肝胰腺中EcCTL的表达量较对照组均显著增加(P<0.05), 且具有明显的时间差异性。本研究证明脊尾白虾C-型凝集素在其免疫反应中起到重要作用, 为进一步探索脊尾白虾免疫系统打下基础。 |
| 关键词: 脊尾白虾(Exopalaemon carinicauda) C-型凝集素 基因克隆 基因表达 |
| DOI：10.11759/hykx/20141231001 |
| 分类号: |
| 基金项目:国家虾产业技术体系 (CARS-47); 山东省泰山产业领军人才工程项目(LNJY2015002); 国家自然科学基金(31472275); 青岛海洋科学与技术国家实验室鳌山科技创新计划项目(2015ASKJ02) |
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| Cloning and expression analysis of C-type lectin gene of Exopalaemon carinicauda |
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WANG You-kun,LIU Ping,LI Ji-tao,LI Jian
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| Abstract: |
| Based on a hemocyte cDNA library of Exopalaemon carinicauda generated in our laboratory, the C-type lectin cDNA of E. carinicauda (named EcCTL) was cloned by rapid amplification of cDNA ends. EcCTL was 1285-bp long, containing an open reading frame of 1041 bp and encoding a mature protein of 346 amino acids, with a molecular mass of 38.56 kDa. Homology analysis revealed that its amino acid sequence was highly conserved, with homologs in other crustaceans. The degree of similarity of EcCTL with cDNA of Palaemon modestus was 91%. EcCTL expression levels in different tissues were analyzed using quantitative real-time PCR. Its expression was detected in all tested tissues, including hemocytes, gill, hepatopancreas, muscle, ovary, intestine, stomach, and eyestalk, with the highest expression level in the hepatopancreas. After challenge with Vibrio anguillarum and white spot syndrome virus, EcCTL expression was upregulated in the hemocytes and hepatopancreas. Our results imply that EcCTL plays an important role in the immune response in prawns and lays a foundation for studies on the E. carinicauda immune system. |
| Key words: Exopalaemon carinicauda C-type lectin gene cloning gene expression |
