Hayashi Shuhei, Aono Rikizo, Hanai Taizo, Mori Hirotada, Kobayashi Takeshi, Honda Hiroyuki
Department of Biotechnology, School of Engineering, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8603, Japan.
J Biosci Bioeng. 2003;95(4):379-83. doi: 10.1016/s1389-1723(03)80071-3.
To investigate the biological mechanism of organic solvent tolerance (OST), DNA microarrays were used to collect and compare the gene expression profiles of normal and organic solvent-tolerant Escherichia coli strains. First, we compared the tolerant-strain OST3410 to its sensitive parent strain JA300 in the absence of organic solvents. Numerous genes showed higher expression levels in OST3410, and Northern analysis was used to confirm the higher expression level of some genes. Next, the gene expression profiles of JA300 and OST3410 exposed to hexane as an organic solvent were investigated and compared with JA300 before exposure to organic solvent. In OST3410 and JA300, 115 and 47 hexane-induced genes were found, respectively. As candidates for genes related to OST, we focused on six genes: cysD, marA, mg1B, tnaA, tnaB and yihM, which were upregulated by hexane in both strains. When these genes were over-expressed on plasmids, only the marA plasmid increased OST activity. It should be noted that we succeeded in finding a gene related to OST activity using only DNA microarray data, without any biochemical or biological knowledge.
为了研究有机溶剂耐受性(OST)的生物学机制,利用DNA微阵列收集并比较了正常和耐有机溶剂的大肠杆菌菌株的基因表达谱。首先,在不存在有机溶剂的情况下,我们将耐受菌株OST3410与其敏感亲本菌株JA300进行了比较。许多基因在OST3410中显示出较高的表达水平,并使用Northern分析来确认某些基因的较高表达水平。接下来,研究了暴露于有机溶剂己烷的JA300和OST3410的基因表达谱,并与未暴露于有机溶剂的JA300进行了比较。在OST3410和JA300中,分别发现了115个和47个己烷诱导基因。作为与OST相关的基因候选,我们重点关注了六个基因:cysD、marA、mg1B、tnaA、tnaB和yihM,这六个基因在两种菌株中均被己烷上调。当这些基因在质粒上过度表达时,只有marA质粒增加了OST活性。应该指出的是,我们仅使用DNA微阵列数据就成功找到了一个与OST活性相关的基因,而无需任何生化或生物学知识。