沙棘H3K9乙酰化修饰全基因组分析

为了绘制沙棘H3K9乙酰化修饰图谱,确定H3K9乙酰化修饰所调控的基因,该实验通过Western blot验证抗体与组蛋白的结合能力和ChIP-seq验证抗体富集效率,获得全基因组范围内沙棘H3K9乙酰化修饰图谱和调控基因。实验结果表明,H3K9ac抗体与复合物具有较强的结合能力。对富集到的DNA片段进行高通量测序,分别获得2.2×10~7和3.6×10~7条原始序列;唯一比对序列广泛分布于沙棘基因组中,并且在结构基因中的两端具有明显的富集。对富集区进行峰的预测结果显示,共预测出1 011个峰;对峰所处部位基因进行功能预测结果发现,H3K9ac对于沙棘细胞代谢和信号转导基因的表达具有重要调控作用。沙棘片段化DNA的富集以及高通量测序结果证明,抗体能够用于研究沙棘的组蛋白修饰类型,并且绘制了沙棘第一张H3K9乙酰化修饰遗传图谱草图,鉴定出沙棘H3K9乙酰化修饰所调控的基因,为今后研究组蛋白修饰对沙棘基因表达的调控方式奠定了基础。

Genome wide Analysis of H3K9ac in Hippophae rhamnoides

In order to make a rough drawing of H3K9ac and identify genes which regulated by H3K9ac modification, provide information for regulation of gene expression in Hippophae rhamnoides, western blot and ChIP seq were carried out. The results showed that the binding capacity of H3K9ac antibody with histone of H. rhamnoides is tightness. High throughput sequencing of enrichmental DNA fragments obtained 2.2×10⁷ and 3.6×10⁷ raw reads. Unique mapped reads were widely distributed in the genome of H. rhamnoides and obviously enriched in both ends of the structural genes. There are 1 011 peaks were identified by predicting the functions of gene enrichment areas, these results showed that H3K9ac played important roles in gene expression of H. rhamnoides. The enrichment and high throughput sequencing of DNA fragments of H. rhamnoides proves that antibodies can be used to study the types of histone modification. The first H3K9ac modification sketches of genetic and the genes regulated by H3K9ac were identified. It is important for investigating genes expression of H. rhamnoides regulated by histone modification.