Detection of bphAa gene expression of Rhodococcus sp. strain RHA1 in soil using a new method of RNA preparation from soil.
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.
Wang
Yong
Y
National Institute for Agro-Environmental Sciences, 3-1-3 Kan-nondai, Tsukuba, Ibaraki 305-8604, Japan.
Shimodaira
Jun
J
Miyasaka
Tomomichi
T
Morimoto
Sho
S
Oomori
Takanori
T
Ogawa
Naoto
N
Fukuda
Masao
M
Fujii
Takeshi
T
D015964:Gene Expression Regulation, Bacterial; D015971:Gene Expression Regulation, Enzymologic; D007506:Iron-Sulfur Proteins / Q000235:genetics*; D008722:Methods; D010105:Oxygenases / Q000235:genetics*; D011078:Polychlorinated Biphenyls / Q000378:metabolism; D021122:Protein Subunits; D012313:RNA / Q000302:isolation & purification*; D012240:Rhodococcus / Q000201:enzymology* / Q000235:genetics; D012987:Soil / Q000032:analysis*; D012988:Soil Microbiology
D007506:Iron-Sulfur Proteins; D021122:Protein Subunits; D012987:Soil; D012313:RNA; D011078:Polychlorinated Biphenyls; D010105:Oxygenases; C074161:biphenyl-2,3-dioxygenase