Construction of a novel Shigella live-vector strain co-expressing CS3 and LTB/STm of enterotoxigenic E.coli

Figure 1 Construction of plasmid pZCS02.

Figure 2 Expression of CS3 in S. flexneri FWL01(pZCS02) (A) and FWL01(pZHY22) (B): Lane 1: Protein marker; lane 2: Extraction of FWL01(pZCS02); lane 3: Extraction of FWL01(A) and FWL01(pZHY22) (B); lane 4: Extraction of E 44815 (A) and FWL01(B); lane 5: Purified CS3 extraction of FWL01(pZCS02); lane 6: purified CS3 extraction of FWL01(pZCS02); lane 7: Purified CS3 fimbriae; lane 8: Extraction of FWL01.

Figure 3 Immunofluorescence microscopy of FWL01(pZCS02) with primary anti-CS3 serum and followed FITC-labeled goat anti-mouse IgG (A) and with primary PBS and followed FITC-labeled goat anti-mouse IgG (B), electron microscopy of fimbrial CS3 on surface of FWL01(pZCS02) (C) and vector strain S. flexneri FWL01 (D).

Figure 4 Expression of fusion protein LTB/STm of FWL01 (pZCS02). A: SDS-PAGE and Western blotting of fusion protein LTB/STm of FWL01 (pZCS02). Lane 1: Protein marker; lane 2: Lysate of the whole cell of FWL01 (pZCS02); lane 3: Lysate of the whole cell of FWL01 (pZCS02); lane 4: Lysate of FWL01; B: Comparison of the expression of LTB/STm between FWL01 (pZCS02) and FWL01 (pZHY22) under the same conditions. Lane 1: Protein marker; lane 2: Lysate of the whole cell of FWL01 (pZCS02); lane 3: Lysate of the whole cell of FWL01 (pZHY22).

Figure 5 Antibody responses to ETEC antigens and Shigella 2a in orally immunized Balb/c mice.