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白斑狗鲛补体成分 C9 基因的分子特征与表达分析。

Molecular characterization and expression analysis of complement component C9 gene in the whitespotted bambooshark, Chiloscyllium plagiosum.

机构信息

State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, No. 22 Hankou Rd, Gulou District, Nanjing 210093, PR China.

出版信息

Fish Shellfish Immunol. 2013 Aug;35(2):599-606. doi: 10.1016/j.fsi.2013.04.042. Epub 2013 May 17.

Abstract

Complement system is known as highly sophisticated immune defense mechanism for antigen recognition as well as effector functions. Activation of the terminal pathway of the complement system leads to the assembly of terminal complement complexes (C5b-9), which induces the characteristic complement-mediated cytolysis. The lytic activity of shark complement involves functional analogues of mammalian C8 and C9. In this article, a full-length cDNA of C9 (CpC9) is identified from cartilaginous species, the whitespotted bambooshark, Chiloscyllium plagiosum by RACE. The CpC9 cDNA is 2263 bp in length, encoding a protein of 603 amino acids, which shares 42% and 43% identity with human and Xenopus C9 respectively. Through sequence alignment and comparative analysis, the CpC9 protein was found well conserved, with the typical modular architecture in TCCs and nearly unanimous cysteine composition from fish to mammal. Phylogenetic analysis places it in a clade with C9 orthologs in higher vertebrate and as a sister taxa to the Xenopus. Expression analysis revealed that CpC9 is constitutively highly expressed in shark liver, with much less or even undetectable expression in other tissues; demonstrating liver is the primary tissue for C9synthesis. To sum up, the structural conservation and distinctive phylogenetics might indicate the potentially vital role of CpC9 in shark immune response, though it remains to be confirmed by further study.

摘要

补体系统被认为是一种高度复杂的免疫防御机制,可识别抗原并发挥效应功能。补体系统末端途径的激活导致末端补体复合物(C5b-9)的组装,从而诱导特征性的补体介导的细胞溶解。鲨鱼补体的溶细胞活性涉及哺乳动物 C8 和 C9 的功能类似物。在本文中,通过 RACE 从软骨鱼类白边鳍鲨(Chiloscyllium plagiosum)中鉴定出全长 C9(CpC9)cDNA。CpC9 cDNA 长 2263bp,编码 603 个氨基酸的蛋白质,分别与人和非洲爪蟾 C9 的同源性为 42%和 43%。通过序列比对和比较分析,发现 CpC9 蛋白高度保守,具有 TCC 中的典型模块结构,并且从鱼类到哺乳动物的半胱氨酸组成几乎一致。系统发育分析将其置于与高等脊椎动物 C9 同源物的一个分支中,并与非洲爪蟾形成姐妹类群。表达分析表明,CpC9 在鲨鱼肝脏中持续高度表达,在其他组织中表达较少甚至无法检测到;表明肝脏是 C9 合成的主要组织。总之,结构保守性和独特的系统发育可能表明 CpC9 在鲨鱼免疫反应中具有潜在的重要作用,但仍需进一步研究来证实。

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