Bacteriocin-producing lactic acid bacteria are commonly used as natural protective cultures.

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Biology Articles » Biotechnology » Detection of the pediocin gene pedA in strains from human faeces by real-time PCR and characterization of Pediococcus acidilactici UVA1 » Figures

Figures

- Detection of the pediocin gene pedA in strains from human faeces by real-time PCR and characterization of Pediococcus acidilactici UVA1

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Figure 1. Molecular weight determination of the bacteriocin produced by strain UVA1. (a) Coomassie stained SDS-PAGE gel. (b) SDS gel overlaid with the indicator strain Listeria ivanovii HPB28. Std: polypeptide molecular weight standard in kDa, F3c: active FPLC-fraction 10-fold concentrated. CFSc: cell-free supernatant of a UVA1 culture, 10-fold concentrated. CFS: cell-free supernatant not concentrated.

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Figure 2. Localization of the pedA gene on the plasmid by curing and Southern blotting. (a) Agarose gel electrophoresis of plasmid DNA. UVA1: P. acidilactici UVA1. UL5: P. acidilactici UL5. bac-: cured derivative of UVA1. M. Supercoiled DNA ladder in kb. ch: chromosomal DNA band. (b) Southern blot DNA hybridization of the DIG labeled pedA-probe with plasmidic DNA from P. acidilactici bac-, UVA1 and UL5. (c) and (d) Agar-well diffusion assay with CFS of cultures of P. acidilactici UVA1 (c) and its cured derivative P. acidilactici bac- (d).

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Figure 3. Transcription analysis of pedA. pedA-reverse transcription-PCR on cDNA from P. acidilactici bac- (1, 2, 3) or UVA1 (7, 8, 9) after 1 h 30, 2 h 30 and 3 h 30 of growth, respectively, and from P. acidilactici DSM 20284^T(4) or UL5 (6) and P. pentosaceus DSM 20336^T (5) after 2 h 30 of growth. 10: water instead of DNA. lm: low molecular weight DNA ladder (in bp). h: Tridye 100-bp DNA ladder (in bp). Expected product size: 100 bp.

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Figure 4. Real-time PCR on faecal DNA samples. (a) Ct values of spiked samples plotted against spiked cell concentration in faecal DNA sample. (b) Ct values obtained for children (C1-C13) and adult (A1-A4) faecal samples. F0: faecal sample free of DNA. w: water instead of DNA. Values represented are means and standard deviations for 3 repetitions.

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