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胞质型磷脂酶A2基因克隆及其表达量随大黄鱼仔稚鱼生长发育的变化

DOI: 10.3969/j.issn.1006-267x.2014.09.053

Keywords: 大黄鱼,磷脂酶A2,克隆,磷脂,分解代谢,生长发育

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Abstract:

利用同源克隆技术和cDNA末端快速扩增(RACE)技术克隆大黄鱼胞质型磷脂酶A2(cPLA2)基因的cDNA全长。此外,应用实时定量PCR法检测不同日龄大黄鱼仔稚鱼cPLA2基因的表达量。序列分析结果表明:cPLA2基因全长2550bp(GenBank登录号:KF006240.1),其中5'端非编码区176bp,开放阅读框2169bp,3'端非编码区205bp,共编码723个氨基酸。系统进化树分析结果表明:克隆所得大黄鱼cPLA2基因与其他鱼类的cPLA2基因亲缘关系较近,与哺乳动物的亲缘关系较远。定量检测结果表明:大黄鱼仔稚鱼cPLA2基因的表达量随日龄的增加为先显著升高(1、3、7日龄vs.15日龄,P<0.05),在15日龄时达到最高值,随后显著下降(15日龄vs.19日龄,P<0.05),之后趋于平稳。由此可知,大黄鱼从仔稚鱼到幼鱼早期的变形过程中,磷脂分解代谢的关键酶cPLA2基因的表达量呈现有规律的变化,这可能对机体保持体内磷脂的动态平衡、维护细胞膜的流动性具有重要的意义。大黄鱼仔稚鱼cPLA2基因表达量的变化趋势在一定程度上可以衡量大黄鱼消化系统的发育情况。

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