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首页> 外文期刊>Journal of bacteriology >Analysis of Protein Synthesis Rates after Initiation of Chromosome Replication in Escherichia coli
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Analysis of Protein Synthesis Rates after Initiation of Chromosome Replication in Escherichia coli

机译：在大肠杆菌中启动染色体复制后的蛋白质合成速率分析

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摘要

The aim of this study was to investigate whether the synthesis rates of some proteins change after the initiation of replication inEscherichia coli. An intR1 strain, in which chromosome replication is under the control of an R1 replicon integrated into an inactivated oriC, was used to synchronize chromosome replication, and the rates of protein synthesis were analyzed by two-dimensional polyacrylamide gel electrophoresis of pulse-labeled proteins. Computerized image analysis was used to search for proteins whose expression levels changed at least threefold after initiation of a single round of chromosome replication, which revealed 7 out of about 1,000 detected proteins. The various synthesis rates of three of these proteins turned out to be caused by unbalanced growth and the synthesis of one protein was suppressed in theintR1 strain. The rates of synthesis of the remaining three could be correlated only to the synchronous initiation of replication. These three proteins were analyzed by peptide mass mapping and appeared to be the products of the dps, gapA, andpyrI genes. Thus, the expression of the vast majority of proteins is not influenced by the state of chromosome replication, and a possible role of the replication-associated expression changes of the three identified proteins in the cell cycle is not clear.
机译：本研究的目的是研究在大肠杆菌中复制开始后某些蛋白质的合成速率是否发生变化。使用 intR1 菌株，其中染色体复制受整合到灭活的 oriC 中的R1复制子的控制，用于同步染色体复制，蛋白质合成速率为脉冲标记蛋白的二维聚丙烯酰胺凝胶电泳分析。计算机图像分析用于搜索在单轮染色体复制启动后表达水平至少改变三倍的蛋白质，该蛋白质揭示了约1,000种检测到的蛋白质中的7种。结果证明这三种蛋白质的各种合成速率是由不均衡的生长引起的，而 intR1 菌株中一种蛋白质的合成受到抑制。其余三个的合成速率只能与复制的同步启动相关。通过肽质谱分析法分析了这三种蛋白质，它们似乎是 dps ， gapA 和 pyrI 基因的产物。因此，绝大多数蛋白质的表达不受染色体复制状态的影响，并且三个细胞周期蛋白的复制相关表达变化在细胞周期中的可能作用尚不清楚。

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