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该研究通过基因克隆的方法获得了虾夷扇贝DNMT1基因(MyDNMT1)的cDNA,全长5 039 bp,其中包括开放阅读框4 761bp,编码1 586个氨基酸,其5’端非编码区为79 bp,3’端非编码区为199 bp。基因相对表达量分析结果显示,MyDNMT1表达起初呈下降趋势,发育至担轮幼虫时期骤然升高,之后再次降低。发育不同时期整体甲基化水平测定结果显示,全基因组甲基化水平最初呈上升趋势,至囊胚期达到最高,随后降低至担轮幼虫时期后再次升高,发育至早期面盘幼虫时期整体甲基化水平再次下降。结合基因相对表达量分析,全基因组甲基化水平动态变化可能是MyDNMT1基因表达与其酶反应之间存在延迟的表现,且推测发育至担轮幼虫后,全基因组甲基化水平的变化与发育过程中出现细胞分化有关。
In this study, the cDNA of DNMT1 gene (MyDNMT1) was obtained by gene cloning. The full-length cDNA of DNMT1 gene (MyDNMT1) was 5 039 bp in length, including 4 761 bp in open reading frame (ORF) encoding 1 586 amino acids and a 79 bp , 3 ’non-coding region of 199 bp. The results of relative gene expression analysis showed that MyDNMT1 expression initially showed a downward trend and developed rapidly until the growth of the larval stage and then decreased again. The results of the whole methylation level at different developmental stages showed that the methylation level of the whole genome initially increased and reached the highest level at the blastocyst stage and then decreased to the level of the larvae at the helical stage and then increased to the level of the early larval stage The methylation level dropped again. According to the analysis of gene relative expression level, the dynamic changes of genome-wide methylation level may be the result of the delay between MyDNMT1 gene expression and its enzyme reaction. It is speculated that the change of methylation level and development process In the emergence of cell differentiation.