一种微小染色质免疫共沉淀方法的建立  

A Mini Chromatin Immunoprecipitation Assay Method

作　　者：唐华容[1] 谭倩[1] 陈方平[1] 谭三勤[1] 张帆[1] 柳林欣[1] 

机构地区：[1]中南大学湘雅医院血液科,湖南长沙410078

出　　处：《激光生物学报》2009年第4期550-555,共6页Acta Laser Biology Sinica

基　　金：国家自然科学基金项目(30570783)

摘　　要：染色质免疫共沉淀(chromatin immunoprecipitation assay,ChIP)是目前研究蛋白与DNA相互作用的强有力的技术之一。然而传统的ChIP实验方法的最大缺陷是要求大量的细胞数,这限制了它应用于少量细胞数的样品。在传统ChIP实验的基础上综合国外文献建立了一种能在少量细胞样品中进行的染色质免疫共沉淀实验,称之为微小染色质免疫共沉淀(miniChIP)。并通过对诱导前后的MEL细胞中表达的β珠蛋白基因簇组蛋白H4乙酰化(acH4)的研究,证实了其可靠性和特异性。结果显示:高敏位点HS2和活跃基因β-maj的启动子区域存在一定的组蛋白H4乙酰化水平,DMSO诱导后显著增加,而不活跃基因Ey的启动子区域则检测到极低水平的组蛋白H4乙酰化,且诱导后无明显变化。Chromatin immunoprecipitation assay （CHIP） provides a powerful tool to analyze protein-DNA interaction. However, one of the drawbacks of current ChIP protocols is its requirement for large cell numbers, which limits the applicability of ChIP to rare cell samples： Together with the literatures, a miniChlP method was established by improving current ChIP method. And its fesibility was confirmed by analyzing H4 acetylation pattern of the β-globin locus in less MEL cells. Results showed that there were a certain level of aeH4 at HS2 and the promoter of β-maj, which is highly active. After treated by DMSO, a dramatic increase of H4 aeetylation at both HS2 and the promoter of β-maj were oh- served, whereas the promoter region of inactive gene （Ey） remains at low level, and no changes after treated by DMSO.

关 键 词：染色质免疫共沉淀(ChIP) 微小染色质免疫共沉淀(miniChIP) 组蛋白H4乙酰化 MEL细胞 

分 类 号：Q243[生物学—细胞生物学] Q939.91