Journal:

Article Title: Evidence for Natural Horizontal Transfer of the pcpB Gene in the Evolution of Polychlorophenol-Degrading Sphingomonads

doi: 10.1128/AEM.68.9.4495-4501.2002

Figure Lengend Snippet: Bacterial strains used in this study for phylogenetic analysis

Article Snippet: However, the pcpB gene was not found to be part of an extensive operon or to be present on the 100-kb endogeneous plasmid of S. chlorophenolicum ATCC 39723 ( 25 ), and despite several trials, we did not find catabolic plasmids in the Kärkölä sphingomonads either (unpublished observations).

Techniques: Isolation

Journal:

Article Title: Evidence for Natural Horizontal Transfer of the pcpB Gene in the Evolution of Polychlorophenol-Degrading Sphingomonads

doi: 10.1128/AEM.68.9.4495-4501.2002

Figure Lengend Snippet: Dot blot DNA hybridization analysis to detect strains carrying the pcpB gene. The genomic DNA (250 ng) was spotted onto a nylon membrane, and the blot was probed for the pcpB gene. Lanes: for row A, 1, S. chlorophenolicum ATCC 39723; 2, S. chlorophenolicum ATCC 33790T; 3, S. chlorophenolicum RA2; 4, S. chlorophenolicum SR3; 5, N. subarcticum KF1T; 6, N. subarcticum KF3; and 7, N. subarcticum NKF1; for row B, 1, K1; 2, K6; 3, K8; 4, K13; 5, K16; 6, K27; 7, K31; and 8, K33; for row C, 1, K39; 2, K40, 3, K44; 4, K66; 5, K74; 6, K76; 7, K101; and 8, K103; and for row D, 1; K104; and 2, K112.

Article Snippet: However, the pcpB gene was not found to be part of an extensive operon or to be present on the 100-kb endogeneous plasmid of S. chlorophenolicum ATCC 39723 ( 25 ), and despite several trials, we did not find catabolic plasmids in the Kärkölä sphingomonads either (unpublished observations).

Techniques: Dot Blot, DNA Hybridization

Journal:

Article Title: Evidence for Natural Horizontal Transfer of the pcpB Gene in the Evolution of Polychlorophenol-Degrading Sphingomonads

doi: 10.1128/AEM.68.9.4495-4501.2002

Figure Lengend Snippet: Southern blot analysis of genomic DNA from chlorophenol-degrading sphingomonads digested with EcoRI. Lane 1, S. chlorophenolicum ATCC 39723; lane 2, N. subarcticum KF1T; lane 3, strain K6; lane 4, strain K16; lane 5, strain K39; lane 6, strain K40; lane 7, strain K74; lane 8, strain K101; and lane 9, Novosphingobium sp. strain MT1. The number of kilobases is given on the left.

Article Snippet: However, the pcpB gene was not found to be part of an extensive operon or to be present on the 100-kb endogeneous plasmid of S. chlorophenolicum ATCC 39723 ( 25 ), and despite several trials, we did not find catabolic plasmids in the Kärkölä sphingomonads either (unpublished observations).

Techniques: Southern Blot

Journal:

Article Title: Evidence for Natural Horizontal Transfer of the pcpB Gene in the Evolution of Polychlorophenol-Degrading Sphingomonads

doi: 10.1128/AEM.68.9.4495-4501.2002

Figure Lengend Snippet: Western blot analysis used to detect expression of PCP-4-monooxygenase with polyclonal antibody. Sample 1, S. chlorophenolicum ATCC 39723; sample 2, N. subarcticum KF1T, sample 3, strain K16, and sample 4, strain K101. Duplicate samples were incubated with (+) and without (−) 2,3,4,6-TeCP. kDa, kilodaltons.

Article Snippet: However, the pcpB gene was not found to be part of an extensive operon or to be present on the 100-kb endogeneous plasmid of S. chlorophenolicum ATCC 39723 ( 25 ), and despite several trials, we did not find catabolic plasmids in the Kärkölä sphingomonads either (unpublished observations).

Techniques: Western Blot, Expressing, Incubation

Journal:

Article Title: Evidence for Natural Horizontal Transfer of the pcpB Gene in the Evolution of Polychlorophenol-Degrading Sphingomonads

doi: 10.1128/AEM.68.9.4495-4501.2002

Figure Lengend Snippet: Alignment of the predicted amino acid sequences for the S. chlorophenolicum ATCC 39723 (EMBL accession no. M98557) and Novosphingobium sp. strain MT1 (AJ319678) pcpB homologs. The amino acids that are located in the flavin adenine dinucleotide-binding domains and conserved in these two sequences and hydroxybenzoate p-hydroxylase are underlined.

Article Snippet: However, the pcpB gene was not found to be part of an extensive operon or to be present on the 100-kb endogeneous plasmid of S. chlorophenolicum ATCC 39723 ( 25 ), and despite several trials, we did not find catabolic plasmids in the Kärkölä sphingomonads either (unpublished observations).

Techniques: Binding Assay