Song Xiaoyan, Wang Lingling, Song Linsheng, Zhao Jianmin, Zhang Huan, Zheng Peilin, Qiu Limei, Liu Xiaolin, Wu Longtao
College of Animal Science and Technology, Northwest A&F University, Yangling, Shaanxi, 712100, China.
Mol Biol Rep. 2009 Jul;36(6):1637-45. doi: 10.1007/s11033-008-9363-8. Epub 2008 Sep 26.
Cyclophilin A (CypA), a receptor for the immunosuppressive agent cyclosporin A (CsA), is a cis-trans peptidyl-prolyl isomerase (PPIase) which accelerates the cis-trans isomerization of prolyl-peptide bonds, interacts with a variety of proteins and therefore regulates their activities. One CypA (designated CfCypA) cDNA was cloned from Chlamys farreri by expressed sequence tag (EST) and rapid amplification of cDNA ends (RACE) techniques. The full-length cDNA of CfCypA consisted of 1,248 nucleotides with a canonical polyadenylation signal sequence AATAAA, a poly (A) tail, and an open reading frame (ORF) of 495 nucleotides encoding a polypeptide of 164 amino acids. The deduced amino acid sequence shared high similarity with CypA from the other species, indicating that CfCypA should be a new member of the CypA family. Quantitative real-time (RT) PCR was employed to assess the mRNA expression of CfCypA in various tissues and its temporal expression in haemocytes and gonad of scallops challenged with Vibrio anguillarum. The mRNA transcripts of CfCypA could be detected in all the examined tissues with highest expression level in gonad. After bacterial challenge, the expression level of CfCypA was almost unchanged in haemocytes, but up-regulated in gonad and increased to the peak (22.59-fold; P < 0.05) at 4 h post-injection, and then dropped to the original level at 8 h post-injection. These results indicated that CfCypA was constitutive expressed in haemocytes, but could be induced in gonad, and perhaps played a critical role in response to the bacterial challenge in gonad.
亲环蛋白A（CypA）是免疫抑制剂环孢素A（CsA）的受体，是一种顺反肽脯氨酰异构酶（PPIase），可加速脯氨酰肽键的顺反异构化，与多种蛋白质相互作用，从而调节它们的活性。通过表达序列标签（EST）和cDNA末端快速扩增（RACE）技术，从栉孔扇贝中克隆出一个CypA（命名为CfCypA）cDNA。CfCypA的全长cDNA由1248个核苷酸组成，具有典型的多聚腺苷酸化信号序列AATAAA、一个聚（A）尾和一个495个核苷酸的开放阅读框（ORF），编码一个164个氨基酸的多肽。推导的氨基酸序列与其他物种的CypA具有高度相似性，表明CfCypA应该是CypA家族的一个新成员。采用定量实时（RT）PCR评估CfCypA在各种组织中的mRNA表达及其在鳗弧菌攻击的扇贝血细胞和性腺中的时间表达。在所有检测的组织中都能检测到CfCypA的mRNA转录本，性腺中的表达水平最高。细菌攻击后，CfCypA在血细胞中的表达水平几乎没有变化，但在性腺中上调，并在注射后4小时增加到峰值（22.59倍；P<0.05），然后在注射后8小时降至原始水平。这些结果表明，CfCypA在血细胞中组成性表达，但在性腺中可被诱导，可能在性腺对细菌攻击的反应中起关键作用。
