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通过交联反应使(dG-dC)n.(dG-dC)n稳定于Z构象。

Stabilization of (dG-dC)n.(dG-dC)n in the Z conformation by a crosslinking reaction.

作者信息

Castleman H, Hanau L H, Erlanger B F

出版信息

Nucleic Acids Res. 1983 Dec 10;11(23):8421-9. doi: 10.1093/nar/11.23.8421.

Abstract

(dG-dC)n.(dG-dC) was converted to the Z conformer by heating in the presence of Mn++n. Reaction of this preparation with the crosslinking reagent, DL-diepoxybutane (DEB), stabilized this conformer so that it retained its structure even when returned to conditions that favored reversion to the B conformation. Treatment of the crosslinked Z conformer with periodate caused scission of the crosslink, allowing reversion to the B conformer. Reaction of (dG-dC)n.(dG-dC)n in the B conformation with DEB did not prevent conversion to the Z conformer in 4M NaC1; dialysis of the high salt solution against low ionic strength buffer allowed return to the B conformer. The Z in equilibrium B transitions were followed by circular dichroism studies and immunochemical procedures. The results suggest the feasibility of stabilizing Z sequences of DNA in chromatin by crosslinking, so that they could then be identified after DNA isolation.

摘要

在Mn++存在的情况下加热,(dG-dC)n.(dG-dC)被转化为Z构象体。该制剂与交联剂DL-二环氧丁烷(DEB)反应,使这种构象体得以稳定,以至于即使回到有利于恢复为B构象的条件下,它仍能保持其结构。用高碘酸盐处理交联的Z构象体会导致交联断裂,从而使其恢复为B构象体。处于B构象的(dG-dC)n.(dG-dC)n与DEB反应并不妨碍在4M NaCl中转化为Z构象体;将高盐溶液对低离子强度缓冲液进行透析可使其恢复为B构象体。通过圆二色性研究和免疫化学方法跟踪Z与平衡态B之间的转变。结果表明通过交联稳定染色质中DNA的Z序列是可行的,这样在DNA分离后就可以识别它们。

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