文章摘要
王俊丽,秦超彬,卢荣华,雒燕婷,梁丽娜,刘燕琴,聂国兴..淇河鲫甘露糖受体基因克隆及嗜水气单胞菌感染对其基因表达的影响[J].水产学报,2018,42(1):18~28
淇河鲫甘露糖受体基因克隆及嗜水气单胞菌感染对其基因表达的影响
Molecular cloning of mannose receptor in Carassius auratus var. Qihe and effects of A. hydrophila infection on its gene expression
投稿时间:2017-01-24  修订日期:2017-05-28
DOI:10.11964/jfc.20170110699
中文关键词: 淇河鲫  甘露糖受体  基因克隆  基因表达  嗜水气单胞菌
英文关键词: Carassius auratus var. Qihe  mannose receptor  gene cloning  gene expression  Aeromonas hydrophila
基金项目:国家自然科学基金(31372545);河南省高校科技创新团队支持计划(14IRTSTHN013);河南省自然科学基金(162300410165);河南省科技计划基金项目(142300410159);河南省教育厅科学技术重点研究项目(14A180007)
作者单位E-mail
王俊丽 河南师范大学生命科学学院 wangjunli1971@126.com 
秦超彬 河南师范大学水产学院河南省水产动物养殖工程技术研究中心  
卢荣华 河南师范大学水产学院河南省水产动物养殖工程技术研究中心  
雒燕婷 河南师范大学生命科学学院  
梁丽娜 河南师范大学生命科学学院  
刘燕琴 河南师范大学生命科学学院  
聂国兴. 河南师范大学水产学院
河南省水产动物养殖工程技术研究中心 
niegx@htu.cn 
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中文摘要:
      为了解淇河鲫(Carassius auratus var. Qihe)甘露糖受体(Mannose Receptor, MR)的结构特点及其在抗感染免疫反应中的作用,本研究通过同源克隆和RACE技术,获得了淇河鲫甘露糖受体(CaMR)全长cDNA,并采用实时荧光定量PCR(qPCR)技术研究了嗜水气单胞菌感染对CaMR组织表达的影响。结果表明,CaMR cDNA全长4473 bp,5’非编码区81 bp,3’非编码区90 bp,编码一个由1433 aa组成的前体蛋白,其中前20 aa为信号肽。CaMR的氨基酸序列与分子结构与其他物种高度相似:胞外一个富含半胱氨酸的结构域(CRD)、一个纤连蛋白II型结构域(FNIID)及8个串连的C 型凝集素样结构域(CTLDs),一个跨膜区和一个很短的胞内区。氨基酸同源性和进化分析表明,CaMR与草鱼、团头鲂、斑马鱼等鲤科鱼类的进化关系较近,与草鱼MR的同源性最高(82.4%)。通过qPCR检测到头肾、脾、心脏、肌肉等10种组织中均有表达,其中头肾表达量最高;嗜水气单胞菌感染后,头肾、脾、心脏和肌肉的表达量均呈先升后降的趋势,而肠道则呈下降趋势。本研究为进一步揭示CaMR在免疫反应中的作用机制及在淇河鲫疾病防治中的应用奠定了基础。
英文摘要:
      Mannose receptor (MR), a member of the C type lectin receptor superfamily, is thought to be involved in the non-specific immune responses in animals. To illustrate the MR molecular structure of Carassius auratus var. Qihe (CaMR) and the role it played in anti-infection immunity, homology cloning and RACE techniques were used in the present study to obtain the complete cDNA sequence, and the real-time fluorescent quantitative PCR (qPCR) technique was used to measure the effect of Aeromonas hydrophila (A. hydrophila) infection on its gene expression. The full-length cDNA of CaMR contains 4473 bp, including 81bp at 5’UTR and 90 bp at 3’UTR. The open reading frame encodes a putative protein of 1433 aa with a signal peptide of 20 aa. The predicted amino acid sequences showed that CaMR contained a cysteine-rich domain (CRD), a single fibronectin typeⅡ domain (FNⅡD), eight C-type lectin-like domains (CTLDs), a transmembrane domain and a short C-terminal cytoplasmic domain, sharing highly conserved structures with MRs from the other species. Amino acid sequence homology and phylogenic tree showed the close relationship between CaMR and other cyprinid fishes. The MR mRNA could be detected in all the examined tissues with highest level in headkidney. The temporal expression patterns of MR in the headkidney, spleen, heart, and muscle post of infection with A. hydrophila were first rise and then fell, while in the intestine was decreased continually. The current research provides a basis for further revealing the function of CaMR in immune reaction and for its application in disease prevention and cure of Carassius auratus var. Qihe.
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