章佳斌;卢晓文;罗少杰;焦钰;邓岳文;

• 1:广东海洋大学水产学院
• 2:广东省珍珠养殖与加工工程技术研究中心

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

Abstract:

Keywords:

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;

参考文献(References)：

• [1] MEEHAN R R. DNA methylation in animal development[J]. Seminars In Cell&Developmental Biology. 2003, 14:53-65.
• [2] RAZIN A. CpG methylation, chromatin structure and gene silencing-a three-way connection[J]. The EMBO Journal. 1998, 17(17):4905-4908.
• [3] AUTRAN D, HUANCA-MAMANI W, VIELLECALZADA J P. Genomic imprinting in plants:the epigenetic version of an Oedipus complex[J]. Current Opinion In Plant Biology. 2005, 8(1):19-25.
• [4]张海燕,赵洪斌,田亚平,等. DNA甲基化检测技术与应用前景[J].标记免疫分析与临床. 2013, 20(5):348-351.
• [5] SUBRAMANIAM D, THOMBRE R, DHAR A, et al.DNA methyltransferases:a novel target for prevention and therapy[J]. Frontiers In Oncology. 2014, 4:80.
• [6] CUI D, XU X. DNA methyltransferases, DNA methylation,and age-associated cognitive function[J]. International Journal of Molecular Sciences. 2018, 19:13155.
• [7] SUZUKI M M, BIRD A. DNA methylation landscapes:provocative insights from epigenomics[J]. Nature Reviews Genetics. 2008, 9(6):465-476.
• [8] LYKO F, FORET S, KUCHARSKI R, et al. The honey bee epigenomes:differential methylation of brain DNA in queens and workers[J]. Plos Biology. 2010, 8(11):e1000506.
• [9] RIVIERE G, WU G, FELLOUS A, et al. DNA methylation is crucial for the early development in the oyster C. gigas[J]. Marine Biotechnology. 2013, 15(6):739-753.
• [10]罗少杰,张鹏飞,焦钰,等.马氏珠母贝近交与杂交家系的DNA甲基化多态性比较[J].广东海洋大学学报. 2016, 36(6):9-15.
• [11]罗少杰,邓岳文,郑哲,等.马氏珠母贝外套膜不同区域基因组DNA甲基化MSAP分析[J].中国水产科学. 2016, 23(6):1227-1235.
• [12] SONG K, LI L, ZHANG G. The association between DNA methylation and exon expression in the Pacific oyster Crassostrea gigas[J]. Plos One. 2017, 12(9):e185224.
• [13] FARIAS N D, DE OLIVEIRA N F P, DA SILVA P M.Perkinsus infection is associated with alterations in the level of global DNA methylation of gills and gastrointestinal tract of the oyster Crassostrea gasar[J].Journal of Invertebrate Pathology. 2017, 149:76-81.
• [14] DU X, FAN G, JIAO Y, et al. The pearl oyster Pinctada fucata martensii genome and multi-omic analyses provide insights into biomineralization[J]. Gigascience.2017, 6:1-12.
• [15]苏玉,王溪,朱卫国. DNA甲基转移酶的表达调控及主要生物学功能[J].遗传. 2009, 31(11):1087-1093.
• [16]王志刚,建新. DNA甲基转移酶分类、功能及其研究进展[J].遗传. 2009, 31(9):903-912.
• [17] ROUNTREE M R, BACHMAN K E, BAYLIN S B.DNMT1 binds HDAC2 and a new co-repressor, DMAP1,to form a complex at replication foci[J]. Nature Genetics.2000, 25(3):269-277.
• [18] FARISA S, FAGAN R L, MATTHEW W, et al. The replication focus targeting sequence(RFTS)domain is a DNA-competitive inhibitor of Dnmt1[J]. Journal of Biological Chemistry. 2011, 286(17):15344-15351.
• [19] CROSS S H, MEEHAN R R, NAN X, et al. A component of the transcriptional repressor Me CP1shares a motif with DNA methyltransferase and HRX proteins[J]. Nature Genetics. 1997, 16(3):256-259.
• [20] FUNABARA D, OHMORI F, KINOSHITA S, et al.Novel genes participating in the formation of prismatic and nacreous layers in the pearl oyster as revealed by their tissue distribution and rna interference knockdown[J]. Plos One. 2014, 9:e847061.
• [21] RAI K, NADAULD L D, CHIDESTER S, et al. Zebra fish Dnmt1 and Suv39h1 regulate organ-specific terminal differentiation during development[J].Molecular and Cellular Biology. 2006, 26(19):7077-7085.
• [22] ANDERSON R M, BOSCH J A, GOLL M G, et al. Loss of Dnmt1 catalytic activity reveals multiple roles for DNA methylation during pancreas development and regeneration[J]. Developmental Biology. 2009, 334(1):213-223.