A survey of the cellular responses in Pseudomonas putida KT2440 growing in sterilized soil by microarray analysis

作者：Wang Yong; Morimoto Sho; Ogawa Naoto; Fujii Takeshi^*

来源：FEMS Microbiology Ecology, 2011, 78(2): 220-232.

DOI：10.1111/j.1574-6941.2011.01146.x

• gene expression
• RNA extraction
• quantitative RT-PCR
• mRNA amplification

Genome-wide scanning of gene expression by microarray techniques was successfully performed on RNA extracted from sterilized soil inoculated with Pseudomonas putida KT2440/pSL1, which contains a chloroaromatic degrading plasmid, in the presence or absence of 3-chlorobenzoic acid (3CB). The genes showing significant changes in their expression in both the triplicate-microarray analysis using amplified RNA and the single-microarray analysis using unamplified RNA were investigated. Pathway analysis revealed that the benzoate degradation pathway underwent the most significant changes following treatment with 3CB. Analysis based on categorization of differentially expressed genes against 3CB revealed new findings about the cellular responses of the bacteria to 3CB. Genes specifically involved in the transport of 3CB were upregulated, including a K(+)/H(+) antiporter complex, a universal stress protein, two cytochrome P450 proteins and an efflux transporter. The downregulated expression of several genes involved in carbon metabolism and the genes belonging to a prophage in the presence of 3CB was observed. This study demonstrated the applicability of the method of soil RNA extraction for microarray analysis of gene expression in bacteria growing in sterilized soil.

• 出版日期2011-11