SEARCH

SEARCH BY CITATION

Fig. S1. Quantification of the expression rates of the pha genes by real-time RT-PCR assays. Gene transcription profiles of pha genes throughout growth of P. putida KT2442 (A–F), KT42C1 (G–L) and KT42D (M–Q) in M63 0.1 N plus 15 mM sodium octanoate (black bars) or 20 mM glucose (red bars) are shown; samples along the growth curve were taken at 0, 3.5, 7 and 23 h. The gene transcription of phaC1 (A, G, M), phaZ (B, H, N), phaC2 (C, I, O), phaD (D, J), phaF (E, K, P) and phaI (F, L, Q) genes is depicted. An initial concentration of 5 ng cDNA was used for quantitative RT-PCR. Error bars represent standard deviation calculated from the results of three independent experiments. y-axis: transcription level (cDNA ng); x-axis: time of culture (h).

Fig. S2. Gel retardation analyses of PhaD binding to the PI, PC1 and PF promoter regions.

A. The DNA probes used (PI, PC1 and PF) are indicated schematically on the right. Cell extract and gel retardation analyses were performed as described under Experimental procedures. Lanes 2–7 contained 0.086, 0.17, 0.34, 0.69, 1.7 and 3.4 μg of total protein of PhaD+ extracts obtained from cells bearing plasmid pIZD (phaD) respectively. Lane 1 contained no extract and lane 8 contains 3.4 μg of total protein of PhaD-free extracts obtained from cells bearing control plasmid pIZ1016 respectively. The DNA–PhaD complexes are indicated.

B. PhaD binding competition analysis to PI promoter with unlabelled DNA-specific probe (lanes 3, 4, 5) and unrelated salmon sperm DNA (lanes 6, 7, 8). Gel retardation was performed in the presence of 1.7 μg of total protein of PhaD+ extracts. Lane 1 contains no extract.

Fig. S3. Homology-based 3D model of dimeric PhaD. A view of the most representative predicted polar interactions between HOCoA and PhaD. Colour scheme is CPK, with the exception of carbon atoms from the protein (green) or from the ligand (magenta). Oval highlights the 3-hydroxyoctanoyl moiety.

Please note: Wiley-Blackwell are not responsible for the content or functionality of any supporting materials supplied by the authors. Any queries (other than missing material) should be directed to the corresponding author for the article.

FilenameFormatSizeDescription
EMI_2199_sm_Fig_S1_01.tif266KSupporting info item
EMI_2199_sm_Fig_S1_02.tif262KSupporting info item
EMI_2199_sm_Fig_S1_03.tif260KSupporting info item
EMI_2199_sm_Fig_S1_04.tif264KSupporting info item
EMI_2199_sm_Fig_S1_05.tif270KSupporting info item
EMI_2199_sm_Fig_S1_06.tif269KSupporting info item
EMI_2199_sm_Fig_S1_07.tif254KSupporting info item
EMI_2199_sm_Fig_S1_08.tif255KSupporting info item
EMI_2199_sm_Fig_S1_09.tif258KSupporting info item
EMI_2199_sm_Fig_S1_10.tif255KSupporting info item
EMI_2199_sm_Fig_S1_11.tif279KSupporting info item
EMI_2199_sm_Fig_S1_12.tif261KSupporting info item
EMI_2199_sm_Fig_S1_13.tif274KSupporting info item
EMI_2199_sm_Fig_S1_14.tif266KSupporting info item
EMI_2199_sm_Fig_S1_15.tif269KSupporting info item
EMI_2199_sm_Fig_S1_16.tif284KSupporting info item
EMI_2199_sm_Fig_S2_01.tif441KSupporting info item
EMI_2199_sm_Fig_S2_02.tif225KSupporting info item
EMI_2199_sm_Fig_S3.tif364KSupporting info item

Please note: Wiley Blackwell is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.