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两种β防御素基因在团头鲂(Megalobrama amblycephala)中的分子克隆与表达分析。

Molecular cloning and expression analysis of two β-defensin genes in the blunt snout bream (Megalobrama amblycephala).

机构信息

Key Laboratory of Freshwater Animal Breeding, Ministry of Agriculture, College of Fisheries, Huazhong Agricultural University, Wuhan 430070, PR China.

出版信息

Comp Biochem Physiol B Biochem Mol Biol. 2013 Sep;166(1):91-8. doi: 10.1016/j.cbpb.2013.07.006. Epub 2013 Jul 20.

Abstract

β-Defensins are a group of cysteine-rich, cationic antimicrobial peptides that play important roles in innate immune system against pathogenic microbes invading. In this study, the part-length cDNA sequences of two β-defensin genes (maΒD-1, maΒD-2) in blunt snout bream (Megalobrama amblycephala) were identified. Homology analysis showed that the cDNA sequences of maΒD-1 and maΒD-2 had high similarities to those in common carp and zebrafish. Real-time quantitative PCR results exhibited that expression level of maΒD-1 in juvenile tissues was the highest in skin, followed by blood and liver, whereas maΒD-2 was lowly expressed in liver, kidney, brain and foregut. In the early development period, fertilized eggs to 31-day post-hatching (dph) larvae, the expression levels of maΒD-1 were higher at the stage from heart beat stage to 3 dph with the highest value at 1 dph, whereas maΒD-2 was expressed higher at fertilized eggs and late cleavage stages. Following bacterial stimulation in vivo by Aeromonas sobria, maΒD-2 expressions were significantly up-regulated in liver, skin, gill, and foregut of juveniles, and maΒD-1 expressions were significantly up-regulated in liver and skin. The results suggest that maΒD-1 and maΒD-2 may play important roles in protecting blunt snout bream embryos, fry and juveniles from pathogenic microbe invading.

摘要

β-防御素是一组富含半胱氨酸的阳离子抗菌肽,在先天免疫系统抵御入侵的病原微生物中发挥重要作用。本研究鉴定了团头鲂(Megalobrama amblycephala)中两个β-防御素基因(maβD-1、maβD-2)的部分 cDNA 序列。同源性分析表明,maβD-1 和 maβD-2 的 cDNA 序列与鲤鱼和斑马鱼的高度相似。实时定量 PCR 结果显示,maβD-1 在幼鱼组织中的表达水平在皮肤中最高,其次是血液和肝脏,而 maβD-2 在肝脏、肾脏、大脑和前肠中低表达。在早期发育阶段,从受精卵到 31 日龄(dph)幼虫,maβD-1 的表达水平在心跳阶段到 3 dph 较高,1 dph 时最高,而 maβD-2 在受精卵和晚期卵裂阶段表达较高。在体内受到气单胞菌刺激后,幼鱼的肝脏、皮肤、鳃和前肠中 maβD-2 的表达显著上调,肝脏和皮肤中 maβD-1 的表达显著上调。这些结果表明,maβD-1 和 maβD-2 可能在保护团头鲂胚胎、鱼苗和幼鱼免受病原微生物入侵方面发挥重要作用。

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