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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。
Kuntiz type serine proteinase inhibitor (KuSPI), also known as pancreatic trypsin inhibitor, can interact with the corresponding protease to regulate the cascade reaction in vivo. In this study, a total length of 624 bp of KuSPI cDNA was obtained by RACE cloning and contained 38 bp and 148 bp 5 ’and 3’ UTRs. The ORF (open reading frame) 438 bp encoded 145 amino acids with a molecular mass of 16 397.5 u, KuSPI amino acid sequence homology analysis found that it contains a typical Kuntiz domain, is a typical KuSPI gene. qRT-PCR showed that the KuSPI gene was expressed in the caecal muscle, mantle, intestine, axillary, liver, blood, gill, gonad and kidney, and highest in mantle. After induced by Aeromonas hydrophila, the expression level of KuSPI gene in mantle changed most obviously, which showed a significant increase first and then decreased. Mantle membrane is the first line of defense of innate immunity of Hyriopsis cumingii, we speculate that Ku SPI gene plays an important role in its immune response.