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Figure 2 | Microbial Cell Factories

Figure 2

From: Expression of an endotoxin-free S-layer/allergen fusion protein in gram-positive Bacillus subtilis 1012 for the potential application as vaccines for immunotherapy of atopic allergy

Figure 2

Intra- and extracelluar expression of the S-layer/allergen fusion protein rSbpA/Bet v1 by B. subtilis 1012. (A) Extracellular expression. Immunoblot analysis using the polyclonal rabbit antiserum raised against the S-layer protein SbpA. Purified S-layer fusion protein rSbpA/Bet v1 isolated from E. coli BL21Star(DE3) used as positive control (lane 1), SDS-extracted whole cells of B. subtilis 1012 containing plasmid pHT43/sbpA/bet v1 before (lane 2) and 1 h, 2 h and 3 h after induction of fusion protein expression (lane 3, 4 and 5). Supernatant fraction of the B. subtilis 1012 cell culture before (lane 6) and 1 h, 2 h, 3 h after induction of expression (lane 7, 8 and 9). Immunoblot analysis using the monoclonal antibody BIP1 raised against Bet v1. SDS-extracted whole cells of B. subtilis 1012 containing plasmid pHT43/sbpA/bet v1 before (lane 10) and 2 h after induction of fusion protein expression (lane 11). Supernatant fraction of the B. subtilis 1012 cell culture before (lane 12) and 2 h after induction of expression (lane 13). WC: whole cells, SN: supernatant (B) Intracellular expression. Immunoblot analysis using the polyclonal rabbit antiserum raised against the S-layer protein SbpA. SDS-extracted whole cells of B. subtilis 1012 containing plasmid pHT01/sbpA/bet v1 before (lane 1) and 1, 2, 3 h after induction of fusion protein expression (lane 2, 3 and 4). Immunoblot analysis using the monoclonal antibody BIP1 raised against Bet v1. SDS-extracted whole cells of B. subtilis 1012 containing plasmid pHT01/sbpA/bet v1 before (lane 5) and 2 h after induction of fusion protein expression (lane 6).

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