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利用一种从土壤中制备RNA的新方法检测红球菌属菌株RHA1的bphAa基因在土壤中的表达。

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

作者信息

Wang Yong, Shimodaira Jun, Miyasaka Tomomichi, Morimoto Sho, Oomori Takanori, Ogawa Naoto, Fukuda Masao, Fujii Takeshi

机构信息

National Institute for Agro-Environmental Sciences, 3-1-3 Kan-nondai, Tsukuba, Ibaraki 305-8604, Japan.

出版信息

Biosci Biotechnol Biochem. 2008 Mar;72(3):694-701. doi: 10.1271/bbb.70493. Epub 2008 Mar 7.

Abstract

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.

摘要

为了解土壤细菌对周围环境的反应,有必要检测土壤中细菌的基因表达谱。为此,我们开发了一种可重复地从土壤中提取RNA的新方法。使用这种新方法,我们从经过灭菌处理并接种了红球菌属菌株RHA1的田间土壤中提取了RNA,RHA1是一种从γ-六氯环己烷污染土壤中分离出的联苯降解菌。琼脂糖凝胶电泳数据表明提取的RNA得到了适当纯化。这种新方法可轻松应用于大量RNA的制备。通过TaqMan方法进行的实时逆转录-聚合酶链反应(RT-PCR)实验表明,该菌株中参与联苯降解的bphAa基因在添加联苯的土壤中被诱导表达。

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