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将一种DNA甲基转移酶导入果蝇体内以探究体内染色质结构。

Introduction of a DNA methyltransferase into Drosophila to probe chromatin structure in vivo.

作者信息

Wines D R, Talbert P B, Clark D V, Henikoff S

机构信息

Howard Hughes Medical Institute, Basic Sciences Division, Fred Hutchinson Cancer Research Center, Seattle, WA 98104, USA.

出版信息

Chromosoma. 1996;104(5):332-40. doi: 10.1007/BF00337221.

Abstract

The dam DNA methyltransferase gene from Escherichia coli was introduced into Drosophila in order to probe chromatin structure in vivo. Expression of the gene caused no visible defects or developmental delay even at high levels of active methylase. About half of each target site was found to be methylated in vivo, apparently reflecting a general property of chromatin packaged in nucleosomes. Although site-specific differences were detected, most euchromatic and heterochromatic sites showed comparable degrees of methylation, at least at high methylase levels. Methylase accessibility of a lacZ reporter gene subject to position-effect variegation throughout development was only slightly reduced, consistent with studies of chromatin accessibility in vitro. Silencing of lacZ during development differed from silencing of an adjacent white eye pigment reporter gene in the adult, as though chromatin structure can undergo dynamic alterations during development.

摘要

为了在体内探测染色质结构,将来自大肠杆菌的dam DNA甲基转移酶基因导入果蝇。即使在活性甲基化酶水平很高时,该基因的表达也未导致明显缺陷或发育延迟。每个靶位点约一半在体内被甲基化,这显然反映了包装在核小体中的染色质的一般特性。尽管检测到了位点特异性差异,但大多数常染色质和异染色质位点显示出相当程度的甲基化,至少在甲基化酶水平较高时如此。在整个发育过程中受到位置效应斑驳影响的lacZ报告基因的甲基化酶可及性仅略有降低,这与体外染色质可及性研究一致。发育过程中lacZ的沉默与成虫中相邻白眼色素报告基因的沉默不同,好像染色质结构在发育过程中会发生动态变化。

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