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Fig. 4 | Microbial Cell Factories

Fig. 4

From: Protective effects of a food-grade recombinant Lactobacillus plantarum with surface displayed AMA1 and EtMIC2 proteins of Eimeria tenella in broiler chickens

Fig. 4

a The expression cassette for single- or dual-anchored proteins. The promoter PsppA mediated single-anchoring of the mCherryAT in pLQa-pgsA′-mCherry. Each of the co-anchoring expression plasmids (pLQa-p′m-ES and pLQa-ES-p′m) contains the inducible promoter PsppA, followed by two connected single-anchoring cassettes, which were separated by the SD sequence (AGGAAACAGACC). The difference between pLQa-p′m-ES and pLQa-ES-p′m is the splicing order of each anchoring cassette. b Western blot of EGFP using mouse anti-EGFP monoclonal antibody. c Quantitative analysis of western blot shown in b. d Western blot of mCherry using mouse anti-6 × His tag monoclonal antibody. e Quantitative analysis of western blot shown in d. f Representative images of flow cytometric analysis of L. plantarum NC8/Δalr strains containing the plasmids designed for single- or dual-anchoring of EGFP or mCherry. The strains are denoted by different colors in the flow cytometry histograms: pLQa-EGFP-S (green), pLQa-pgsA’-mCherry (red), pLQa-p’m-E (purple), pLQa-ES-p’m (blue). EGFP-A represents the EGFP-expressing strains; mCherry-A represents the mCherry-expressing strains. The L. plantarum NC8/Δalr harboring the empty pLQa-pgsA’ plasmid was used as a negative control (gray). (h) Statistical analysis of the flow cytometry results obtained by one-way ANOVA (NS: no significant difference, P > 0.05, ***P < 0.001)

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