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Figure S1. Effect of pigX and pigZ mutations on Serratia ATCC 39006 prodigiosin production (A) and pigB gene expression (B). Samples (1 ml) for prodigiosin assays and pigB qRT-PCR analysis were taken from early stationary phase (10 h) cultures of a pigX mutant (CH9), a pigZ mutant (TG39) and a pigX, pigZ double mutant (SP15), grown in LB broth. Prodigiosin was measured as absorbance at 534 nm, as described previously (Fineran et al., 2005b). Quantitative PCR was performed on the pigB gene, as representative of the entire prodigiosin operon, as described previously (Williamson et al., 2008). A 5 µl aliquot of template DNA was used in a 25 µl reaction containing 1× SYBR Green PCR Master Mix (Applied Biosystems) and 10 pmol of primers for pigB (Table S1). PCR amplification was performed using an Applied Biosystems Prism 7300 sequence detection system, using reaction conditions as described previously (Williamson et al., 2008). Fluorescence data were processed using SDS software (ABI) to produce threshold cycle (Ct) values for each sample. Relative gene expression was obtained using 16S rRNA as the control with mRNA/16S rRNA = 1 in the wild-type control. Data shown are the means ± SD of three independent experiments.

Table S1. Bacterial strains, phages and primers used in this study.

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FilenameFormatSizeDescription
EMI4_140_sm_FigS1.tif339KSupporting info item
EMI4_140_sm_TabS1.doc30KSupporting 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.