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. 2017 Aug 22;12(8):e0183346. doi: 10.1371/journal.pone.0183346

Fig 4. Increased atxA copy number enhances pagA gene transcription and PA content in the B. anthracis secretome.

Fig 4

(A) Genetic structure of pPAGK plasmid. TT—T4 phage transcription terminator. (B) Active transcription of atxA and pagA in BH490 derivatives containing the pPAGK plasmid. Two-step qPCR results of atxA and pagA transcription respectively from cultures grown in NBY broth with 0.8% NaHCO3 in 15% CO2 (top panels) and in air (bottom panels). Relative expression represents the 2-ΔΔCt (RQ) value normalized to Ames 35 and evaluated with three reference genes: rpoB, gyrB, and dnaJ. Values reported are the mean relative expression ± standard error of the mean calculated from ExpressionSuite and GraphPad Prism software. An asterisk indicates a significant difference (p<0.05) from an unpaired t-test compared to Ames 35. (C) Western blot analysis of PA production in strains grown in the same conditions as in panel B.