Affinity purification of MLL3/MLL4 histone H3K4 methyltransferase complex.

Affinity purification of MLL3/MLL4 histone H3K4 methyltransferase complex.
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DOI:
10.1007/978-1-61779-376-9_30
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发表时间:
2012
期刊:
Methods in molecular biology (Clifton, N.J.)
影响因子:
--
通讯作者:
Ge, Kai
Ge, Kai
中科院分区:
其他
文献类型:
--
作者:
Cho, Young-Wook;Hong, SunHwa;Ge, Kai

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组蛋白H3赖氨酸4 (H3K4)甲基化与活性转录基因相关。在哺乳动物细胞中,存在多个set1样组蛋白H3K4甲基转移酶复合物,它们具有重叠但不同的亚基组成。开发分离这些组蛋白H3K4甲基转移酶复合物的方法将有助于了解组蛋白H3K4甲基化调节哺乳动物基因表达的分子机制。在本章中,我们使用flag标记的PA1 (MLL3/MLL4复合物的独特亚基)提供了一种一步亲和纯化方案,用于分离MLL3/MLL4组蛋白H3K4甲基转移酶复合物。
Methylation on histone H3 lysine 4 (H3K4) correlates with actively transcribed genes. In mammalian cells, there exist multiple Set1-like histone H3K4 methyltransferase complexes, which have overlapping but distinct subunit compositions. Developing methods to isolate each of these histone H3K4 methyltransferase complexes would help understand the molecular mechanisms by which histone H3K4 methylation regulates mammalian gene expression. In this chapter, we provide a one-step affinity purification protocol on isolation of the MLL3/MLL4 histone H3K4 methyltransferase complex using FLAG-tagged PA1, a unique subunit of the MLL3/MLL4 complex.
DOI: 10.1073/pnas.0707292104
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