Ferraro A, Eufemi M, Cervoni L, Altieri F, Turano C
Department of Biochemical Sciences A. Rossi Fanelli, University La Sapienza, Rome, Italy.
Acta Biochim Pol. 1995;42(2):145-51.
To detect the interactions of DNA with the nuclear matrix proteins, DNA-protein cross-linkages were induced in intact nuclei from chicken liver by the use of cis-diammine dichloroplatinum. Methods have been devised for fast purification both of the proteins and of the DNA fragments involved in the cross-linked complexes. By Southern-Western blotting a number of matrix proteins isolated from the complexes have been shown to recognize specifically DNA sequences present in the cross-linked DNA fragments. This experimental approach not only allows to identify the nuclear matrix-DNA interactions existing in the nucleus before its disruption, but also provides a preparation of matrix proteins enriched in those species which are involved in such interactions and which can therefore be detected with high sensitivity.
为了检测DNA与核基质蛋白的相互作用，利用顺二氯二氨铂在鸡肝完整细胞核中诱导DNA-蛋白质交联。已经设计出了快速纯化参与交联复合物的蛋白质和DNA片段的方法。通过Southern-Western印迹法，从复合物中分离出的许多核基质蛋白已被证明能特异性识别交联DNA片段中存在的DNA序列。这种实验方法不仅能够识别在细胞核破坏之前存在于细胞核中的核基质与DNA的相互作用，而且还能制备富含参与此类相互作用的蛋白质的核基质蛋白制剂，因此能够以高灵敏度检测到这些蛋白质。
