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Figure 5 | Microbial Cell Factories

Figure 5

From: Improved adhesive properties of recombinant bifidobacteria expressing the Bifidobacterium bifidum-specific lipoprotein BopA

Figure 5

Adhesion of B. bifidum S17 in competition to purified BopA-His 6 . SDS-PAGE (A) and Western Blot (B) for detection of BopA-His6 in crude extract of E. coli BL21 (DE3) pMGS_bopA His6 before (lane 1) or after (lane 2) induction with 0.013 mM L-arabinose. Lane 3: purified BopA-His6 protein in pooled and dialysed fractions of the Ni-NTA-affinity chromatography. M: protein ladder. (C) Adhesion of B. bifidum S17 to T84 (black bars), Caco-2 (light grey bars) and HT29 (dark grey bars) cells after pre-incubation with 1, 10 or 100 μg/ml BopA-His6. As control IECs were incubated with 100 μg/ml BSA instead of BopA-His6. Adherent bacteria were determined after growth on MRSC agar plates. Results are shown as percent adherent bacteria relative to the positive control (untreated cells). Values are mean ± standard deviation of one representative of at least three independent experiments performed in triplicate measurements. Statistical analysis was performed by pairwise comparison to the respective positive control (i.e. adhesion of B. bifidum S17 without pre-incubation with protein) using Students t-test. Letters above bars indicate statistical significance (a: p < 0.05; b: p < 0.01; c: p < 0.001).

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