Kashiwagi Akiko, Sakurai Takahiro, Tsuru Saburo, Ying Bei-Wen, Mori Kotaro, Yomo Tetsuya
Department of Bioinformatic Engineering, Graduate School of Information Science and Technology, Osaka University, 2-1 Yamadaoka, Suita, Osaka 565-0871, Japan.
Metab Eng. 2009 Jan;11(1):56-63. doi: 10.1016/j.ymben.2008.08.002. Epub 2008 Aug 22.
We generated 61 strains of Escherichia coli in which the expression level of a specific single gene can be changed continuously over a physiologically significant range. In each strain, one auxotrophic gene was deleted from its original position and reinserted at a specific position on the chromosome under the control of the tetA promoter. Therefore, the level of expression of the target gene can be controlled easily by altering the concentrations of inducers, e.g., anhydrotetracycline and doxycycline, in the medium. Protein and mRNA levels and changes in proliferation rate were examined in some of the strains in our collection to determine the ability to control the level of target gene expression over a physiologically significant range. These strains will be useful for extracting omics data sets and for the construction of genome-scale mathematical models, because causality between perturbations in gene expression level and their consequences can be clearly determined.
我们构建了61株大肠杆菌菌株，在这些菌株中，特定单基因的表达水平能够在具有生理意义的范围内连续变化。在每一株菌株中，一个营养缺陷型基因从其原始位置被删除，并在染色体上tetA启动子的控制下重新插入到一个特定位置。因此，通过改变培养基中诱导剂（如脱水四环素和强力霉素）的浓度，可轻松控制目标基因的表达水平。我们检测了部分菌株的蛋白质和mRNA水平以及增殖速率的变化，以确定在具有生理意义的范围内控制目标基因表达水平的能力。这些菌株将有助于提取组学数据集以及构建基因组规模的数学模型，因为基因表达水平的扰动与其后果之间的因果关系能够被明确确定。
