马氏珠母贝DNA甲基化转移酶Dnmt1的克隆及表达Molecular Cloning and Expression Analysis of DNA Methyltransferase 1 (Dnmt1) from Pinctada martensii
章佳斌;卢晓文;罗少杰;焦钰;邓岳文;
摘要(Abstract):
【目的】对马氏珠母贝(Pinctada martensii)DNA甲基化转移酶1(DNA methyltransferase 1,Dnmt1)的基因全长及组织表达作分析,探究马氏珠母贝甲基转移酶Dnmt1(PmDnmt1)参与贝壳矿化调控机理。【方法】利用RACE技术获得PmDnmt1的c DNA序列全长,并对PmDnmt1的序列特征进行分析,同时利用逆转录-聚合酶链反应RT-PCR技术检测马氏珠母贝不同组织中PmDnmt1的表达情况。【结果与结论】PmDnmt1基因c DNA长4 818 bp,其中,开放阅读框为4 623 bp,编码1 540个氨基酸。预测分子质量约为174.55 ku、理论等电点为5.89。结构域分析显示,PmDnmt1具有DMAP结合结构域、DNMT1-RFD结构域、锌指结构、BAH结构域和C5胞嘧啶DNA甲基化酶结构域等。多序列比对结果发现,Dnmt1在不同物种间具有较高保守性。实时荧光定量PCR分析显示PmDnmt1在所有检测组织中均有表达,在外套膜中央膜部分的表达量最高(P <0.05)。PmDnmt1可通过调控外套膜的DNA甲基化修饰参与贝壳的形成。
关键词(KeyWords): 马氏珠母贝;DNA甲基化转移酶;基因克隆;表达模式
基金项目(Foundation): 国家自然科学基金(31672626)
作者(Authors): 章佳斌;卢晓文;罗少杰;焦钰;邓岳文;
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