This work was supported by a grant from The National Natural Sciences Foundation of China (30200158).
染色质结构和基因表达调节是当前国际前沿研究的热点.染色质免疫沉淀法是研究染色质结构的首选方法,它不仅可用来研究体内反式因子与DNA的相互作用,也可以用来研究组蛋白修饰与基因表达的关系.综合国外相关文献,建立了一种简便的染色质免疫沉淀法,并通过对诱导前后的MEL细胞中β-珠蛋白基因簇组蛋白H3乙酰化的研究,证实了其可操作性.结果表明:高敏位点HS2和活跃基因βmaj的启动子区域存在较高的组蛋白乙酰化水平,且诱导前后变化显著,而不活跃基因Ey的启动子区域则几乎检测不到组蛋白的乙酰化,且诱导前后无明显变化.这一结果与以前的报道相吻合.
Chromatin structure plays a critical role in eukaryotic gene expression. Chromatin immunoprecipitation assay (ChIP) provides a powerful tool to analyze the interaction of trans-acting factors with specific chromatin regions in vivo, as well as the role of histone modifications in gene regulation. A simple ChIP protocol is established and used to study the H3 acetylation pattern of the β-globin locus in MEL cells. DMSO induction results in a dramatic increase in H3 acetylation at hypersensitive site HS2 and active gene (βmaj) promoter, whereas the inactive gene (Ey) promoter maintains low acetylation. This result indicates that the ChIP method is feasible.
傅湘辉,刘德培,辛立,郝德龙,董文吉,黄粤,梁植权.一种简便的染色质免疫沉淀法的建立[J].生物化学与生物物理进展,2003,30(4):634-638
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