Table 1.
Bacterial strains, plasmids and oligonucleodide primers used in this study.
| Description | Contents | Reference/source |
|---|---|---|
| Strains | ||
| E. coli TG1 | supE thi-1 Δ(lac-proAB) Δ(mcrB-hsdSM)5(rK- mK-) [F' traD36 proAB lacIqZΔM15 | Stratagene |
| B. subtilis 1012 | leuA8, metB5, hsrM1, nonA | CatchMaps BV, MoBiTec |
| Plasmids | ||
| pET28a/sbpA/bet v1 | Cloning of rsbpA/bet v1 | [15] |
| pHT01 | Ampr, Cmr, Pgrac promoter (consisting of groEpromotor, the lacO operator and the gsiBSD sequence, ColE1 ori,, lacI repressor, E. coli/B. subtilis shuttle vector | MoBiTec |
| pHT43 | see pHT01 + amyQ signal sequence | MoBiTec |
| pHT01/sbpA/bet v1 | Expression of rSbpA/Bet v1 in cytoplasm of B. subtilis 1012 | This study |
| pHT43/sbpA/bet v1 | Expression of rSbpA/Bet v1 in cytoplasm of B. subtilis 1012 followed by subsequent secretion into the culture medium | This study |
| Primers* | ||
| SbpA/AatII/forward | 5'-cgcgacgtcgcgcaagtaaacgactataacaaatc-3' | This study |
| Bet v1/SmaI/reverse | 5'-tcccccgggttagttgtaggcatcggagtgtg-3' | This study |
*Bold nucleotide sequences indicate restriction endonuclease sites.