Stoichiometric analysis of protein- and nucleic acid-based structures in the cell nucleus
Stoichiometric analysis of protein- and nucleic acid-based structures in the cell nucleus
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DOI：
10.1016/s0968-4328(99)00019-0
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发表时间：
1999-04-01
期刊：
影响因子：
2.4
通讯作者：
Kruhlak, MJ
中科院分区：
文献类型：
--
作者：
Bazett-Jones, DP;Hendzel, MJ;Kruhlak, MJ
我们描述了一种方法来选择性地成像的蛋白质为基础的架构在真核细胞核中使用氮和磷映射。此外，我们描述了一种方法来确定总质量以及蛋白质和RNA之间的化学计量关系。这种方法是用细胞核中称为染色质间颗粒的颗粒结构来说明的。在这样做时，我们表明，这些颗粒含有异质核RNA，并具有平均蛋白质和RNA含量分别为3.094和1.672 MDa。我们还测试了磷检测的灵敏度外源应用纯化的双链DNA的薄切片的表面，并已表明，结构小到单分子的双链DNA可以在原位检测使用这些电子光谱成像技术。(C)1999 Elsevier Science Ltd.保留所有权利。
We describe a method to image selectively the protein-based architecture in the eukaryotic cell nucleus using nitrogen and phosphorus mapping. In addition, we describe a method to determine total mass as well as stoichiometric relationships between protein and RNA. This method is illustrated using particulate structures in the nucleus called interchromatin granules. In so doing, we demonstrate that these granules contain heterogeneous nuclear RNA, and have an average protein and RNA content of 3.094 and 1.672 MDa, respectively. We also tested the sensitivity of phosphorus detection by exogenously applying purified duplex DNA to the surfaces of thin sections, and have shown that structures as small as single molecules of duplex DNA can be detected in situ using these electron spectroscopic imaging techniques. (C) 1999 Elsevier Science Ltd. All rights reserved.