杂色鲍(Haliotis diversicolor)硒结合蛋白1基因的克隆及其应激表达

MOLECULAR CLONING AND EXPRESSION OF SELENIUM-BINDING PROTEIN 1 IN HALIOTIS DIVERSICOLOR UNDER STRESSES

硒结合蛋白1(selenium-binding protein 1,SBP1)是一种高度保守的蛋白,广泛参与了机体的多个理化过程。本研究首次获得了杂色鲍(Haliotis diversicolor)SBP1基因的全长c DNA序列2269 bp,并命名为Hd SBP1,ORF为1494 bp,共编码497个氨基酸。荧光定量PCR结果显示,Hd SBP1基因在杂色鲍各组织中均有表达,其中在鳃和肾中的表达量相对较高。缺氧处理后,Hd SBP1在鳃组织和血细胞中的表达量分别在处理24 h和192 h后显著升高(P<0.05);高温应激之后,在鳃组织中,Hd SBP1的表达量在第1时相与第3时相显著升高(P<0.05),而在血细胞中只有在第3时相表现出显著性差异;缺氧和高温联合应激之后,鳃组织中Hd SBP1的表达量在192 h显著上调(P<0.05),在血细胞中Hd SBP1的表达量在0 h、4 h和24 h时均显著升高(P<0.05);副溶血弧菌注射感染之后,在鳃组织中Hd SBP1的表达量在6 h和24 h时显著性上调(P<0.05),而血细胞中Hd SBP1基因的表达量在每个时相的实验组都显著高于对照组(P<0.05)。以上不同的应激条件均会导致Hd SBP1在不同组织中的表达量的显著变化,说明Hd SBP1在杂色鲍的免疫反应中可能扮演着重要的角色。

Selenium-binding protein 1(SBP1) is a highly conserved protein which is widely involved in multiple physical and chemical processes. We cloned a full-length c DNA of SBP1 from Haliotis diversicolor for the first time and named it Hd SBP1. The full-length c DNA sequence is 2269 bp, with an open reading frame of 1494 bp encoding a protein of 497 amino acids. The real-time quantitative PCR results showed that ubiquitous expression of Hd SBP1 in all examined tissues, with highest expression in gills and kidney. Hd SBP1 transcripts were compared under normoxia, and environmental stresses(hypoxia, heat, and Vibrio) in the gills and haemocytes of the abalone. Under hypoxia, the expression of Hd SBP1 m RNA in gills and haemocytes was significantly increased at 24 h and 192 h respectively(P<0.05). Under thermal stress, it was significantly up-regulated in gills at the first phase(when temperature reached 28°C) and the third phase(3 h post thermal stress), so did in haemocytes at the third phase(P<0.05). Under thermal plus hypoxia stress, it was significantly up-regulated in gills at 192 h(P<0.05), and in haemocytes, the expression was significantly up-regulated at 0, 4, and 24 h. After Vibrio parahaemolyticus injection, the expression was significantly up-regulated in gills at 6 h and 24 h and so did in haemocytes at all phases(P<0.05). All these results indicate that Hd SBP1 may play important roles in immunoreaction of H. diversicolor.