Table 1 Bacterial strains, plasmids and oligonucleodide primers used in this study.
Description | Contents | Reference/source |
|---|---|---|
Strains | Â | Â |
E. coli TG1 | supE thi-1 Δ(lac-proAB) Δ(mcrB-hsdSM)5(rK- mK-) [F' traD36 proAB lacIqZΔM15 | Stratagene |
B. subtilis 1012 | leuA 8, metB 5, hsrM 1, nonA | CatchMaps BV, MoBiTec |
Plasmids | Â | Â |
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 v 1 | 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* | Â | Â |
SbpA/AatII/forward | 5'-cgcgacgtc gcgcaagtaaacgactataacaaatc-3' | This study |
Bet v1/SmaI/reverse | 5'-tcccccggg ttagttgtaggcatcggagtgtg-3' | This study |