Novel methylation specific bisulfite primer pairs for epigenetic studies of Capsicum spp.

Ayse Gul Ince, M. Karaca
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Abstract

Over the past ten years, interest in epigenetic has rapidly increased. Heritable and stable changes in gene expression without any change in DNA sequence is in the field of epigenetics. Plants have a well-preserved epigenetic signature called DNA methylation. It is an essential epigenetic mark that protects genomic stability, silences harmful transposon insertions, and controls global gene expression in all developmental stages and environmental circumstances. All three sequence contexts, the asymmetric CpHpH context and the symmetric CpG and CpHpG contexts (where H is C, A, or T), are among DNA methylation sites in plants. Particularly, DNA cytosine methylation affects a wide range of biological processes, such as gene expression, chromatin structure, DNA packing, recombination, genomic imprinting, and DNA replication. The choice of primer pairs that flank cytosine methylation contexts is critical when designing for the detection of DNA cytosine methylation using bisulfite sequencing. We have developed and synthesized 26 bisulfite specific primer pairs suitable for DNA cytosine methylation investigations in peppers. These primers are specific to certain promoters, intergenic regions, and gene bodies (exons, introns, and UTRs). DNA samples taken from various tissues and developmental stages of Capsicum annuum L. Demre Sivrisi were analyzed by these primer pairs to confirm their utilization.
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用于辣椒属植物表观遗传学研究的新型甲基化特异性亚硫酸氢盐引物对
过去十年来,人们对表观遗传学的兴趣迅速增加。在不改变 DNA 序列的情况下,基因表达发生可遗传的稳定变化,这就是表观遗传学的研究领域。植物有一种保存完好的表观遗传特征,叫做 DNA 甲基化。它是一种重要的表观遗传标记,可保护基因组的稳定性,抑制有害的转座子插入,并在所有发育阶段和环境条件下控制全局基因表达。在植物的 DNA 甲基化位点中,有三种序列上下文,即不对称的 CpHpH 上下文和对称的 CpG 和 CpHpG 上下文(其中 H 是 C、A 或 T)。特别是,DNA胞嘧啶甲基化会影响一系列生物过程,如基因表达、染色质结构、DNA包装、重组、基因组印记和DNA复制。在设计使用亚硫酸氢盐测序法检测 DNA 胞嘧啶甲基化时,选择侧翼胞嘧啶甲基化上下文的引物对至关重要。我们开发并合成了 26 对亚硫酸氢盐特异引物,适用于辣椒 DNA 胞嘧啶甲基化研究。这些引物对某些启动子、基因间区和基因体(外显子、内含子和 UTR)具有特异性。我们用这些引物对辣椒不同组织和不同发育阶段的 DNA 样本进行了分析,以确认它们的用途。
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8 weeks
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