摘要
【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cDNA 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cDNA 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。
【Objective】In order to offer a theoretical basis for exploring the regulation of DNA methyltransferase 1 of Pinctada martensii(PmDnmt1) in biomineralization, the full-length gene was obtained and the tissue expression pattern of PmDnmt1 was elucidated.【Method】The cDNA sequence of PmDnmt1 was obtained by rapid amplification of cDNA ends technology(RACE).The characteristics of PmDnmt1 were analyzed, and the mRNA expression of PmDnmt1 in different tissues of P.martensii were detected by fluorescence quantitative PCR.【Result and conclusion】The results showed that the length of PmDnmt1 was 4 818 bp,including ORF 4 623 bp which encoded 1 540 amino acids.The predicted molecular weight was about 174.55 ku and the theoretical isoelectric point was 5.89.The result of domain prediction analysis showed that PmDnmt1 had DMAP binding domain, Dnmt1 RFD, zinc finger structure, BAH domain and c-5 cytosine-specific DNA methylase domain profile.The multi-sequence alignment results showed that Dnmt1 was highly conserved across different species.Real-time fluorescence quantitative PCR analysis showed that PmDnmt1 gene was expressed in all test tissue, with the highest expression in the mantle central(P<0.05).PmDnmt1 may be involved in shell formation through regulating the DNA methylation.
作者
章佳斌
卢晓文
罗少杰
焦钰
邓岳文
ZHANG Jia-bin;LU Xiao-wen;LUO Shao-jie;JIAO Yu;DENG Yue-wen(Fisheries College of Guangdong Ocean University, Zhanjiang 524088, China;Pearl Breeding and Processing Engineering Technology Research Centerof Guangdong Province, Zhanjiang 524025, China)
出处
《广东海洋大学学报》
CAS
2019年第5期16-23,共8页
Journal of Guangdong Ocean University
基金
国家自然科学基金(31672626)
