Construction and characterization of bivalent vaccine candidate expressing HspA and Mr18000 OMP from Helicobacter pylori

Figure 1 Schematic construction of plasmid pET32a(+)/HspA-Omp₁₈.

Figure 2 10 g·L^-1 agarose gel electrophoresis of HspA DNA fragment amplified by PCR from Helicobacter pylori. Lane 1. PCR marker, Lane 2. Negative control, Lane 3. PCR products.

Figure 3 The identification of recombinant vector by PCR. Lane 1. DNA Marker, Lane 2, 3. PCR products with the template of Top10/recombinant vector, and recombinant vector respectively, Lane 4. Negative control.

Figure 4 Identification of recombinant plasmid by restriction enzyme digestion. Lane 1. pET32a(+)/HspA digested by BamHI, Lane 2. pET32a(+)/HspA-Omp₁₈ digested by BamHI, Lane 3. DNA marker, Lane 4. pET32a(+)/HspA-Omp₁₈ digested by BamHI and Hind III simultaneously, Lane 5. pET32a(+)/ HspA-Omp₁₈ digested by kpnI, BamHI and Hind III simultaneously.

Figure 5 150 g·L^-1 SDS-PAGE of total protein in recombinant vector expressed in BL21 E. coli. Lane 1. Standard protein marker (M_r 14; 31; 42; 66; 97 × 10³), Lane 2. Bacterial protein expressed in BL21 after induction for 4 hours with IPTG, Lane 3-7. Ex-pression of recombinant vector in BL21 after induction for 4 h with IPTG.

Figure 6 Antigenic analysis of the expression of recombinant vector by Western blot. Lane 1. BL21/pET32a(+)/HspA-Omp₁₈, Lane 2. BL21/pET32a(+), Lane 3. BL21/pET32a(+)/HspA.