4.4 Article

Detection of bphAa gene expression of Rhodococcus sp strain RHA1 in soil using a new method of RNA preparation from soil

Journal

BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY
Volume 72, Issue 3, Pages 694-701

Publisher

TAYLOR & FRANCIS LTD
DOI: 10.1271/bbb.70493

Keywords

soil; RNA; real-time reverse transcription-polymerase chain reaction (RT-PCR); Rhodococcus; biphenyl

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To understand the response of soil bacteria to the surrounding environment, it is necessary to examine the gene expression profiles of the bacteria in the soil. For this purpose, we developed a new method of extracting RNA from soil reproducibly. Using this new method, we extracted RNA from a field soil, which was sterilized and inoculated with Rhodococcus sp. strain RHA1, a biphenyl degrader isolated from gamma-hexachlorocyclohexane-contaminated soil. Data from agarose gel electrophoresis indicated that the extracted RNA was purified properly. This new method can be applied easily in the preparation of large amounts of RNA. Real-time reverse transcription-polymerase chain reaction (RT-PCR) experiments performed by the TaqMan method suggested that the bphAa gene in this strain, which is involved in the degradation of biphenyl, was induced in the biphenyl amended soil.

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