Keasling J D, Palsson B O, Cooper S
Department of Microbiology and Immunology, University of Michigan Medical School, Ann Arbor 48109-0620.
J Bacteriol. 1991 Apr;173(8):2673-80. doi: 10.1128/jb.173.8.2673-2680.1991.
F plasmid replication during the Escherichia coli division cycle was investigated by using the membrane-elution technique to produce cells labeled at different times during the division cycle and scintillation counting for quantitative analysis of radioactive plasmid DNA. The F plasmid replicated, like the minichromosome, during a restricted portion of the bacterial division cycle; i.e., F plasmid replication is cell-cycle specific. The F plasmid replicated at a different time during the division cycle than a minichromosome present in the same cell. F plasmid replication coincided with doubling in the rate of enzyme synthesis from a plasmid-encoded gene. When the cell cycle age of replication of the F plasmid was determined over a range of growth rates, the cell size at which the F plasmid replicated followed the same rules as did replication of the bacterial chromosome--initiation occurred when a constant mass per origin was achieved--except that the initiation mass per origin for the F plasmid was different from that for the chromosome origin. In contrast, the high-copy mini-R6K plasmid replicated throughout the division cycle.
利用膜洗脱技术在大肠杆菌分裂周期的不同时间标记细胞,并通过闪烁计数对放射性质粒DNA进行定量分析,从而研究了F质粒在大肠杆菌分裂周期中的复制情况。F质粒与微型染色体一样,在细菌分裂周期的特定时间段内进行复制;也就是说,F质粒的复制具有细胞周期特异性。在分裂周期中,F质粒的复制时间与同一细胞中存在的微型染色体不同。F质粒的复制与质粒编码基因的酶合成速率加倍同时发生。当在一系列生长速率范围内确定F质粒复制的细胞周期年龄时,F质粒复制时的细胞大小遵循与细菌染色体复制相同的规则——当每个复制起点达到恒定质量时开始复制——只是F质粒每个复制起点的起始质量与染色体复制起点的不同。相比之下,高拷贝的mini-R6K质粒在整个分裂周期中都进行复制。