Direct imaging of DNA in living cells reveals the dynamics of chromosome formation.
Direct imaging of DNA in living cells reveals the dynamics of chromosome formation.
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DOI：
10.1083/jcb.144.5.813
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发表时间：
1999-03-08
期刊：
影响因子：
--
通讯作者：
Cook PR
中科院分区：
文献类型：
--
作者：
Manders EM;Kimura H;Cook PR
在大多数体细胞的间期核中，不能直接看到个别的染色体；它们只能在有丝分裂时才能看到。我们已经开发出一种方法，可以在活细胞中直接观察DNA链，并用它来分析有丝分裂染色体是如何形成的。将天然前体三磷酸胸苷的荧光类似物(例如，Cy5-dUTP)引入细胞，然后在共聚焦显微镜的加热舞台上生长。这种类似物被内源性酶结合到DNA中。由于识别和去除不寻常残基的机制不会阻止细胞周期的后续进展，现在的荧光DNA链可以在组装成染色体并分离到女儿和孙子的过程中被跟随。活细胞中这种链的电影表明，在G2期，染色体轴沿着简单可识别的路径穿过它们的领土，而后期复制区域在前期染色体形成时保持其相对位置。定量分析证实，在染色体凝聚的这一阶段，个别区域几乎没有移动。因此，间期染色体区域的大体结构与前期染色体的大体结构直接相关。
Individual chromosomes are not directly visible within the interphase nuclei of most somatic cells; they can only be seen during mitosis. We have developed a method that allows DNA strands to be observed directly in living cells, and we use it to analyze how mitotic chromosomes form. A fluorescent analogue (e.g., Cy5-dUTP) of the natural precursor, thymidine triphosphate, is introduced into cells, which are then grown on the heated stage of a confocal microscope. The analogue is incorporated by the endogenous enzymes into DNA. As the mechanisms for recognizing and removing the unusual residues do not prevent subsequent progress around the cell cycle, the now fluorescent DNA strands can be followed as they assemble into chromosomes, and segregate to daughters and granddaughters. Movies of such strands in living cells suggest that chromosome axes follow simple recognizable paths through their territories during G2 phase, and that late replicating regions maintain their relative positions as prophase chromosomes form. Quantitative analysis confirms that individual regions move little during this stage of chromosome condensation. As a result, the gross structure of an interphase chromosome territory is directly related to that of the prophase chromosome.
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影响因子：
7.8
作者：
Minden, J S;Agard, D A;Sedat, J W;Alberts, B M
通讯作者：
Alberts, B M
DOI：
10.1101/sqb.1993.058.01.085
发表时间：
1993-01-01
期刊：
COLD SPRING HARBOR SYMPOSIA ON QUANTITATIVE BIOLOGY
影响因子：
--
作者：
CREMER, T;KURZ, A;LICHTER, P
通讯作者：
LICHTER, P
影响因子：
64.8
作者：
HIRAOKA, Y;MINDEN, JS;AGARD, DA
通讯作者：
AGARD, DA
影响因子：
64.8
作者：
Heald, R;Tournebize, R;Karsenti, E
通讯作者：
Karsenti, E
影响因子：
56.9
作者：
CHALFIE, M;TU, Y;PRASHER, DC
通讯作者：
PRASHER, DC