| Estimation of the Chromosomal Copy Number in Synechococcus elongatus PCC 7942
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Author:
Date:
2016-07-05
[Abstract] Cyanobacteria are prokaryotic organisms that perform oxygenic photosynthesis. Freshwater cyanobacteria, such as Synechococcus elongatus PCC 7942 and Synechocystis sp. PCC 6803, are model organisms for the study of photosynthesis, gene regulation, and biotechnological applications because they are easy to manipulate genetically. However, while studying these cyanobacteria, care has to be taken with respect to genetic heterogeneity in the establishment of gene disruptants, because these cyanobacteria contain multiple chromosomal copies per cell. Here, we describe a method for the estimation of chromosomal copy number in Synechococcus 7942. Using this method, we have recently observed that the chromosomal copy number of Synechococcus 7942 ...
[摘要] 蓝细菌是进行含氧光合作用的原核生物。 淡水蓝细菌，例如细长聚球藻PCC 7942和聚集细胞 PCC 6803是用于研究光合作用，基因调节和生物技术应用的模式生物，因为它们易于基因操作。 然而，当研究这些蓝细菌时，必须注意遗传异质性在基因破坏物的建立中，因为这些蓝细菌每个细胞含有多个染色体拷贝。 在这里，我们描述了用于估计在聚球藻7942中的染色体拷贝数的方法。使用这种方法，我们最近观察到，聚球藻7942的染色体拷贝数显着地改变 其生长阶段。 这种技术可用于研究多倍体不仅在蓝细菌，而且在其他多倍体生物。
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| Chromatin Fractionation Assay in Fission Yeast
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Author:
Date:
2014-07-20
[Abstract] The protein recruitment onto chromatin is a critical process for DNA metabolism, including DNA replication, DNA repair and DNA recombination. Especially DNA modification enzymes and checkpoint proteins are loaded onto DNA damage sites in a context-dependent manner. In our recent study (Kunoh and Habu, 2014), the chromatin association of Pcf1, a large subunit of Chromatin Assembly Factor-1 (CAF-1), was monitored after exposure of cells to hydroxyurea which slowed down the DNA replication. Results of the chromatin fractionation assay provided evidence that Pcf1 was recruited to chromatin upon DNA replication stress. A similar procedure enabled to reveal the chromatin association of Orp1, Mcm proteins, and Swi6 (Sadaie et al., 2008; Ogawa et al., 1999). This assay allows us ...
[摘要] 染色质上的蛋白质募集是DNA代谢的关键过程，包括DNA复制，DNA修复和DNA重组。特别是DNA修饰酶和检查点蛋白以上下文依赖性方式装载到DNA损伤位点。在我们最近的研究中（Kunoh和Habu，2014），在将细胞暴露于羟基脲（其减慢DNA复制）后，监测Pcf1的染色质缔合，其为染色质装配因子-1（CAF-1）的大亚基。染色质分馏测定的结果提供了Pcf1在DNA复制应激时募集到染色质的证据。类似的程序能够揭示Orp1，Mcm蛋白和Swi6的染色质缔合（Sadaie等人，2008; Ogawa等人，1999）。该测定允许我们从活细胞分离染色质结合和非结合蛋白。以下各部分的免疫印迹提供了关于我们的靶蛋白的染色质结合状态的信息。
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