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在染色体中分离与特定核蛋白相互作用的DNA序列的通用方法:高迁移率族蛋白HMG-T与鱼精蛋白基因家族一个亚组的结合

General method for isolation of DNA sequences that interact with specific nuclear proteins in chromosomes: binding of the high mobility group protein HMG-T to a subset of the protamine gene family.

作者信息

Blanco J, States J C, Dixon G H

出版信息

Biochemistry. 1985 Dec 31;24(27):8021-8. doi: 10.1021/bi00348a028.

Abstract

A general method is described for the isolation of the DNA with which specific nuclear proteins interact in chromosomes. This method is based on the covalent photo-cross-linking of nuclear proteins to the DNA sequences, to which they normally bind, by means of irradiation with UV light and the selective retrieval of specific subsets of protein-DNA adducts by using specific antibodies. The application of this procedure to isolate the DNA sequences with which the trout high mobility group protein (HMG-T) interacts has shown that in trout liver this protein associates specifically with DNA sequences in proximity to a subset of the family of protamine genes but not with the histone or vitellogenin genes. From these observations, it appears that the HMG-T protein may be associated with inactive gene sequences.

摘要

本文描述了一种分离与染色体中特定核蛋白相互作用的DNA的通用方法。该方法基于通过紫外线照射使核蛋白与它们通常结合的DNA序列进行共价光交联,并使用特异性抗体选择性地回收蛋白质-DNA加合物的特定子集。将该程序应用于分离与鳟鱼高迁移率族蛋白(HMG-T)相互作用的DNA序列,结果表明在鳟鱼肝脏中,该蛋白与一组精蛋白基因家族附近的DNA序列特异性结合,但不与组蛋白或卵黄蛋白原基因结合。从这些观察结果来看,HMG-T蛋白可能与无活性基因序列相关。

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