Good Start Genetics, Inc., 237 Putnam Ave. Cambridge, MA 02139, USA.
Methods. 2012 Nov;58(3):212-20. doi: 10.1016/j.ymeth.2012.06.017. Epub 2012 Jul 6.
Bacterial chromosomes must be compacted by three-orders of magnitude to fit within the cell. While such compaction could in theory yield disordered structures, it is becoming increasingly clear that bacterial chromosomes are in fact arranged in regular and reproducible fashions and that their configurations are tightly connected to fundamental processes such as chromosome segregation. Nonetheless, due to throughput and resolution limitations associated with traditional assays, many question regarding bacterial chromosome structure and its relation to genome function remain. Here, I review the related technologies, chromosome conformation capture (3C) and chromosome conformation capture carbon copy (5C), which my collaborators and I recently introduced as tools to probe the high-resolution folding of entire bacterial genomes. These technologies utilize covalent cross-linking and proximity ligation to facilitate the measurement of the spatial positioning of hundreds of genomic loci, thereby opening the door to high-throughput studies of bacterial chromosome structure. Hence, 3C and 5C represent powerful new tools for assaying the three-dimensional architecture of bacterial genomes.
细菌染色体必须经过三个数量级的压缩才能容纳在细胞内。虽然这种压缩在理论上可能产生无序结构，但越来越明显的是，细菌染色体实际上是以规则且可重复的方式排列的，并且它们的构型与染色体分离等基本过程紧密相关。尽管如此，由于传统检测方法的通量和分辨率限制，许多关于细菌染色体结构及其与基因组功能关系的问题仍然存在。在这里，我回顾了相关技术，即染色体构象捕获（3C）和染色体构象捕获碳拷贝（5C），这是我的合作者和我最近引入的工具，用于探测整个细菌基因组的高分辨率折叠。这些技术利用共价交联和邻近连接来促进数百个基因组位点的空间定位测量，从而为细菌染色体结构的高通量研究打开了大门。因此，3C 和 5C 代表了用于分析细菌基因组三维结构的强大新工具。
