三角帆蚌KuSPI基因的克隆及表达分析

发布时间：2018-11-11 14:53

【摘要】：Kuntiz型丝氨酸蛋白酶抑制剂(Kuntiz-type serine proteinase inhibitor,KuSPI)又称胰腺胰蛋白酶抑制剂,能够和相应的蛋白酶互作来调控生物体内的级联反应。本研究通过RACE克隆得到三角帆蚌KuSPI cDNA全序列共624 bp,包含38 bp和148 bp的5'和3'端UTR,ORF(open reading frame)438 bp,编码氨基酸145个,分子量为16 397.5 u,KuSPI氨基酸序列的同源性分析后发现其包含1个典型Kuntiz结构域,属于典型的KuSPI基因。qRT-PCR检测表明:KuSPI基因在闭壳肌、外套膜、肠、斧足、肝、血液、鳃、性腺和肾脏中均有表达,在外套膜中最高。利用嗜水气单胞菌进行诱导后发现,KuSPI基因在外套膜中表达量变化最大,呈现明显的先升高再降低趋势。外套膜是三角帆蚌先天免疫的首道防线,我们推测Ku SPI基因在其免疫反应中起到重要作用。
[Abstract]:Kuntiz serine protease inhibitor (Kuntiz-type serine proteinase inhibitor,KuSPI), also known as pancreatic trypsin inhibitor, can interact with protease to regulate the cascade reaction in vivo. In this study, the complete KuSPI cDNA sequence of Hyriopsis cumingii was cloned by RACE. The total 624 bp, sequence contained 5 'and 3' -terminal UTR,ORF (open reading frame) 438 bp, encoding 145 amino acids of 38 bp and 148 bp, the molecular weight of which was 16 397.5 u. Homology analysis of KuSPI amino acid sequence revealed that it contains a typical Kuntiz domain, which belongs to a typical KuSPI gene. QRT-PCR analysis showed that KuSPI gene was found in obturator muscle, coat membrane, intestine, axe foot, liver, blood, Gill, and Gill. Both gonads and kidneys were expressed, and the highest expression was found in the mantle. After induction by Aeromonas hydrophila, it was found that the expression of KuSPI gene was the most varied in the coat membrane, showing a tendency of first increasing and then decreasing. The coat membrane is the first line of defense for congenital immunity of Hyriopsis cumingii. We speculate that Ku SPI gene plays an important role in the immune response of Hyriopsis cumingii.
【作者单位】：上海海洋大学农业部淡水水产种质资源重点实验室;上海市水产养殖工程技术研究中心;
【基金】：国家自然科学基金项目(31101939) 上海市教委科研创新重点项目(13ZZ128) 上海高校知识服务平台项目(ZF1206)共同资助
【分类号】：S917.4

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