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Subtractive hybridization and random arbitrarily primed PCR analyses of a benzoate-assimilating bacterium, Desulfotignum balticum

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Subtractive hybridization (SH) and random arbitrarily primed PCR (RAP-PCR) were used to detect genes involved in anaerobic benzoate degradation by Desulfotignum balticum. Through SH, we obtained 121 DNA sequences specific for D. balticum but not for D. phosphitoxidans (a non-benzoate-assimilating species). Furthermore, RAP-PCR analysis showed that a 651-bp DNA fragment, having 55% homology with the solute-binding protein of the ABC transporter system in Methanosarcina barkeri, was expressed when D. balticum was grown on benzoate, but not on pyruvate. By shotgun sequencing of the fosmid clone (38,071 bp) containing the DNA fragment, 33 open reading frames (ORFs) and two incomplete ORFs were annotated, and several genes within this region corresponded to the DNA fragments obtained by SH. An 11.3-kb gene cluster (ORF10–17) revealed through reverse transcription-PCR showed homology with the ABC transporter system and TonB-dependent receptors, both of which are presumably involved in the uptake of siderophore/heme/vitamin B12, and was expressed in response to growth on benzoate.

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This work was carried out as a part of “The Project for Development of Technologies for Analyzing and Controlling the Mechanism of Biodegrading and Processing,” which was supported by the New Energy and Industrial Technology Development Organization (NEDO).

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Correspondence to Hiroshi Habe.

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Habe, H., Kobuna, A., Hosoda, A. et al. Subtractive hybridization and random arbitrarily primed PCR analyses of a benzoate-assimilating bacterium, Desulfotignum balticum . Appl Microbiol Biotechnol 79, 87–95 (2008). https://doi.org/10.1007/s00253-008-1414-5

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  • Sulfate-reducing bacteria
  • ABC transporter
  • Desulfotignum balticum
  • Benzoate assimilation