Release of compact nucleoids with characteristic shapes from Escherichia coli.
Release of compact nucleoids with characteristic shapes from Escherichia coli.
复制标题
从大肠杆菌中释放具有特征形状的紧凑核仁。
DOI：
10.1128/jb.183.17.5041-5049.2001
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发表时间：
2001
影响因子：
3.2
通讯作者：
Murphy,LD
中科院分区：
文献类型：
--
作者：
Zimmerman,SB;Murphy,LD
细菌的基因组DNA包含在一个或几个被称为类核的致密体中。我们描述了一个简单的程序，保留了一般的形状和压实的类核从大肠杆菌后，细胞裂解和类核释放的细胞包膜。该程序是用于制备亚精胺类核（在亚精胺存在下释放的类核）（T。Kornberg，A.洛克伍德和A. Worcel，Proc. Natl. Acad. Sci. USA 71：3189-3193，1974）。加入聚赖氨酸以防止细胞裂解时发生的类核的正常解压缩。在指数期细胞的裂解物中或在用氯霉素或萘啶沙处理以改变类核形态的细胞的裂解物中，类核保留其特征形状。利福平处理的细胞的显着不稳定的类核在这样的裂解物中以紧凑、稳定的形式获得。在多聚赖氨酸存在下释放的类核苷酸容易处理，并提供明确的DNA荧光和相衬图像。在甲醛固定和十二烷基硫酸钠加热后，可以分离出保持特征形状的均匀的类核种群。
The genomic DNA of bacteria is contained in one or a few compact bodies known as nucleoids. We describe a simple procedure that retains the general shape and compaction of nucleoids fromEscherichia coliupon cell lysis and nucleoid release from the cell envelope. The procedure is a modification of that used for the preparation of spermidine nucleoids (nucleoids released in the presence of spermidine) (T. Kornberg, A. Lockwood, and A. Worcel, Proc. Natl. Acad. Sci. USA 71:3189–3193, 1974). Polylysine is added to prevent the normal decompaction of nucleoids which occurs upon cell lysis. Nucleoids retained their characteristic shapes in lysates of exponential-phase cells or in lysates of cells treated with chloramphenicol or nalidixate to alter nucleoid morphology. The notably unstable nucleoids of rifampin-treated cells were obtained in compact, stable form in such lysates. Nucleoids released in the presence of polylysine were easily processed and provided well-defined DNA fluorescence and phase-contrast images. Uniform populations of nucleoids retaining characteristic shapes could be isolated after formaldehyde fixation and heating with sodium dodecyl sulfate.