Molecular and biochemical characterization of the tetralin degradation pathway in Rhodococcus sp. strain TFB.
Bottom Line: In addition to tetralin induction, TFB thn structural genes are subject to glucose repression.Primer extension assays and translational thnA1::gfp and thnS::gfp fusions were used to characterize putative promoter regions.A mutational analysis of the thnA1 promoter region allowed us to define nucleotides within the cis regulatory elements that are important for the control of thn gene expression.
Affiliation: Centro Andaluz de Biología del Desarrollo-CSIC, Universidad Pablo de Olavide Carretera de Utrera, Km 1. 41013-Seville, Spain.Show MeSH
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Mentions: Given that the bph genes in Rhodococcus sp. RHA1 are located on the linear plasmids pRHL1 and pRHL2 (Masai et al., 1997; McLeod et al., 2006), we decided to isolate large plasmids from TFB bacteria. Pulse field electrophoresis analysis revealed two large linear plasmids, denoted pTFB1 and pTFB2, with estimated sizes of 1100 and 280 kb respectively (Fig. 3A). Using a thnD DNA fragment as the probe for southern blotting, we found that the thn genes are located on the pTFB1 plasmid (Fig. 3B).
Affiliation: Centro Andaluz de Biología del Desarrollo-CSIC, Universidad Pablo de Olavide Carretera de Utrera, Km 1. 41013-Seville, Spain.