BACTERIAL-GROWTH CONTROL STUDIED BY FLOW-CYTOMETRY
BACTERIAL-GROWTH CONTROL STUDIED BY FLOW-CYTOMETRY
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DOI：
10.1016/0923-2508(91)90020-b
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发表时间：
1991-02-01
影响因子：
2.6
通讯作者：
LOBNEROLESEN, A
中科院分区：
文献类型：
--
作者：
BOYE, E;LOBNEROLESEN, A
通过采用流式细胞术，可以快速且高精度地确定单个细菌细胞的DNA含量和细胞大小。 此外，在用利福平和细胞分裂抑制剂头孢氨苄一起处理细胞后，可以测量大肠杆菌细胞中的DNA复制起点的数目。 与野生型细胞相反，某些突变体以高频率包含许多不同于2n的起点，表明突变体不会同时在所有起点启动DNA复制。 在这里，我们给出的证据表明，这种damy表型不能发生异常的染色体分离或细胞分裂的结果，但只能引起一个和同一个细胞内的多个起始事件的协调缺陷。 流式细胞术已被用于对大肠杆菌的生长和细胞周期进行准确和详细的分析。杆菌 当细菌培养物的DNA分布保持稳定状态生长时，细胞DNA含量在培养物接近并进入稳定期时降低。 只有在稳定期长时间孵育后，细胞才含有完全复制的染色体，快速生长的细胞在稳定期结束时含有2或4条染色体。
By employing flow cytometry, the DNA content and cell size of individual bacterial cells may be determined rapidly and with high precision. Also, the number of DNA replication origins in Escherichia coli cells can be measured after treating the cells with rifampicin together with the cell division inhibitor cephalexin. As opposed to wild type cells, certain mutants contain, with high frequency, a number of origins different from 2n, indicating that the mutants do not initiate DNA replication at all origins simultaneously. Here we give evidence that this asynchrony phenotype cannot occur as a consequence of aberrant chromosomal segregation or cell division, but can only be caused by defective coordination of multiple initiation events within one and the same cell. Flow cytometry has been used to perform exact and detailed analyses of the growth and cell cycle of E. coli. While the DNA distribution of a bacterial culture was unchanged as long as steady-state growth was maintained, the cellular DNA content was reduced when the culture approached and entered stationary phase. Only after prolonged incubation in stationary phase did the cells contain fully replicated chromosomes, and rapidly growing cells ended up with either 2 or 4 chromosomes in stationary phase.