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从菲律宾帘蛤中克隆和鉴定两种亲环素 A 基因同工型。

Molecular cloning and characterization of two isoforms of cyclophilin A gene from Venerupis philippinarum.

机构信息

Faculty of Life Science and Biotechnology of Ningbo University, Ningbo 315211, PR China.

出版信息

Fish Shellfish Immunol. 2011 Dec;31(6):1218-23. doi: 10.1016/j.fsi.2011.07.001. Epub 2011 Jul 18.

Abstract

Cyclophilin A (CypA) is a ubiquitously distributed intracellular protein belonging to the immunophilin family, which is recognized as the cell receptor for the potent immunosuppressive drug cyclosporine A. In the present study, two isoforms of cyclophilin A gene (named as VpCypA1 and VpCypA2) were isolated and characterized from Venerupis philippinarum by RACE approaches. Both VpCypA1 and VpCypA2 possessed all conserved features critical for the fundamental structure and function of CypA, indicating that the two isoforms of cyclophilin A should be new members of CypA family. The expression of VpCypA2 mRNA in haemocytes was significantly up-regulated and the highest expression level was detected at 96 h post-infection with 7.7-fold increase compared with that in the blank group. On the contrary, the relative expression level of VpCypA1 mRNA was down-regulated rapidly at 6 h post-infection and reached 0.4-fold of the control group. They exhibited different expression profile and identical effect of immune modulation, which might suggest the two VpCypA isoforms exert their function in a manner of synergy. These results provide valuable information for further exploring the roles of cyclophilin A in the immune responses of V. philippinarum.

摘要

亲环素 A(CypA)是一种广泛分布于细胞内的蛋白质,属于免疫亲和素家族,被认为是强效免疫抑制剂环孢素 A 的细胞受体。在本研究中,通过 RACE 方法从菲律宾蛤仔中分离和鉴定了两种亲环素 A 基因(命名为 VpCypA1 和 VpCypA2)。VpCypA1 和 VpCypA2 都具有亲环素 A 的基本结构和功能所必需的所有保守特征,表明这两种亲环素 A 同工型应该是亲环素 A 家族的新成员。VpCypA2 mRNA 在血细胞中的表达水平显著上调,在感染后 96 h 时表达水平最高,与空白组相比增加了 7.7 倍。相反,VpCypA1 mRNA 的相对表达水平在感染后 6 h 时迅速下调,达到对照组的 0.4 倍。它们表现出不同的表达谱和相同的免疫调节作用,这可能表明这两种 VpCypA 同工型以协同的方式发挥作用。这些结果为进一步研究亲环素 A 在菲律宾蛤仔免疫反应中的作用提供了有价值的信息。

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