Table 2.
Strains and expression constructs.
| Strain or plasmid | Description | Source |
|---|---|---|
| Geobacillus stearothermophilus NRS 2004/3a | S-layer glycoprotein-covered neotype strain | N. R. Smith collection[43] |
| E. coli DH5α | F-ϕ80 LacZΔM15 Δ(lacZYA-argF)U169 recA1 endA1 hsdR17(rk−, mk+) phoA supE44 thi-1 gyrA96 relA1 λ− | Invitrogen |
| E. coli BL21 Star(DE3) | F- ompT hsdSB (rB-mB-) gal dcm rne131 (DE3) | Novagen |
| pET28a(+) | E. coli expression vector, used for introducing a C-terminal hexahistidine-tag to the cloned gene; Kmr | Novagen |
| pET30a(+) | E. coli expression vector, used for introducing an N-terminal hexahistidine-tag to the cloned gene; Kmr | Novagen |
| pET28a-A_SgsE(C) | pET-28a(+)/SgsE31−903; pET-28a(+) containing sgsE from G. stearothermophilus NRS 2004/3a devoid of its signal peptide (aa 1-30), C-terminal hexahistidine-tag; Kmr | This study |
| pET28a-C_SgsE(C) | pET-28a(+)/C_SgsE131−903; pET-28a(+) containing sgsE from G. stearothermophilus NRS 2004/3a, devoid of aa 1-100 of the mature S-layer protein; C-terminal hexahistidine-tag; Kmr | This study |
| pET28a-G_SgsE(C) | pET-28a(+)/G_SgsE331−903; pET-28a(+) containing sgsE from G. stearothermophilus NRS 2004/3a, devoid of aa 1-300 of the mature S-layer protein; C-terminal hexahistidine-tag; Kmr | This study |
| pET30a-A_SgsE(N) | pET-30a(+)/SgsE31−903; pET-30a(+) containing sgsE from G. stearothermophilus NRS 2004/3a devoid of its signal peptide (aa 1-30); N-terminal hexahistidine-tag; Kmr | This study |
| pET30a-C_SgsE(N) | pET-30a(+)/SgsE131−903; pET-30a(+) containing sgsE from G. stearothermophilus NRS 2004/3a, devoid of aa 1-100 of the mature S-layer protein; N-terminal hexahistidine-tag; Kmr | This study |
| pET30a-G_SgsE(N) | pET-30a(+)/SgsE331−903; pET-30a(+) containing sgsE from G. stearothermophilus NRS 2004/3a, devoid of aa 1-300 of the mature S-layer protein; N-terminal hexahistidine-tag; Kmr | This study |
| pET28a-A_SgsE | pET-28a(+)/SgsE31−903; pET-28a(+) containing sgsE from G. stearothermophilus NRS 2004/3a, devoid of its signal peptide (amino acids 1-30); Kmr | This study |
| pET28a-C_SgsE | pET-28a(+)/SgsE131−903; pET-28a(+) containing sgsE from G. stearothermophilus NRS 2004/3a, devoid of the first 100 amino acids of the mature S-layer protein; Kmr | This study |
| pET28a-G_SgsE | pET-28a(+)/SgsE331−903; pET-28a(+) containing sgsE from G. stearothermophilus NRS 2004/3a, devoid of the first 300 amino acids of the mature S-layer protein; Kmr | This study |
| pET28a-C_SgsE·RmlA | pET28a-C_SgsE containing RmlA fused to the C-terminus of the S-layer protein; Kmr | This study |
| pET28a-G_SgsE·RmlA | pET28a-G_SgsE containing RmlA fused to the C-terminus of the S-layer protein; Kmr | This study |
| pET28a-RmlA | pET-28a(+) containing rmlA from G. stearothermophilus NRS 2004/3a; C-terminal hexahistidine tag; Kmr | This study |