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从大黄鱼(Pseudosciaena crocea)中克隆和鉴定铁蛋白 H 和 M 亚基。

Gene cloning and characterization of ferritin H and M subunits from large yellow croaker (Pseudosciaena crocea).

机构信息

State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, PR China.

出版信息

Fish Shellfish Immunol. 2010 May-Jun;28(5-6):735-42. doi: 10.1016/j.fsi.2009.11.027. Epub 2009 Dec 22.

Abstract

Ferritin, an iron-binding protein, plays a role in host immune response to invading stimulus. Two ferritin subunit cDNAs, lycfh and lycfm, were identified from large yellow croaker, Pseudosciaena crocea. Analysis of the lycfh and lycfm 5'-UTR showed that both had a putative iron responsive element (IRE). Amino acid sequence alignment and phylogenetic analysis indicated that LycfH and LycfM had high identity to H and M type subunit of vertebrate ferritins respectively, while both of them showed the low identity to L type subunit. In all eight tissues examined, lycfh and lycfm expressions were detectable on transcriptional level. Real-time PCR assay indicated that the transcriptional levels of lycfh and lycfm were significantly increased in liver, spleen and kidney of the fish stimulated by attenuated live Vibrio anguillarum strain. Furthermore, ferritin subunit H and M were probably two candidates of immune molecules involved in acute phase response in large yellow croaker because their transcriptional levels were quickly increased and reached the peak levels at 12 h in liver after challenged with bacteria.

摘要

铁蛋白是一种铁结合蛋白,在宿主对入侵刺激的免疫反应中发挥作用。从大黄鱼(Pseudosciaena crocea)中鉴定出两种铁蛋白亚基 cDNA,即 lycfh 和 lycfm。对 lycfh 和 lycfm 的 5'-UTR 进行分析表明,两者都具有潜在的铁反应元件(IRE)。氨基酸序列比对和系统发育分析表明,LycfH 和 LycfM 分别与脊椎动物铁蛋白的 H 和 M 型亚基具有高度同源性,而它们与 L 型亚基的同源性较低。在所检测的 8 种组织中,lycfh 和 lycfm 的转录水平均可检测到。实时 PCR 检测表明,在受到减毒活鳗弧菌菌株刺激的鱼的肝脏、脾脏和肾脏中,lycfh 和 lycfm 的转录水平显著增加。此外,铁蛋白亚基 H 和 M 可能是大黄鱼急性相反应中涉及的免疫分子的两个候选者,因为它们的转录水平在受到细菌攻击后 12 小时内迅速增加并达到肝脏中的峰值水平。

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