TABLE 3.
Strains and plasmids used in this study
| Strains or plasmids | Relevant properties | Source |
|---|---|---|
| Strains | ||
| E. coli Top10 | Host for propagation of the recombinant plasmids | Invitrogen |
| E. coli BL21-Gold (DE3) | pET system hos strain | Invitrogen |
| S. parasanguinis FW213 | Wild type | |
| S. parasanguinis FW213 Fap1 deletion mutant | Wild type; Fap1 knockout; Fap1::aphA3;Kanr | 8 |
| S. parasanguinis FW213 dGT1 deletion mutant | Wild type; dGT1 knockout; dGT1::aphA3;Kanr | 27 |
| Strains used to produce recombinant proteins | ||
| CgT | pET-sumo: CgT transformed into BL21 | In this study |
| His-CgT | pET-28b: CgT transformed into BL21 | In this study |
| CgTΔH1–3 | pET-sumo: CgTΔH1–3 transformed into BL21 | In this study |
| CgT-DH1 | pET-sumo: CgT-DH1 transformed into BL21 | In this study |
| CgT-DH2 | pET-sumo: CgT-DH2 transformed into BL21 | In this study |
| CgT-DH3 | pET-sumo: CgT-DH3 transformed into BL21 | In this study |
| His-sFap1-GlcNAc-Glc | pET-28b: sFap1 and pvpt-Gtf123 co-transformed into BL21 | 27 |
| His-sFap1-GlcNAc-Glc-Glc | pET-28b: sFap1 and pvpt-Gtf123-DUF1792 co-transformed into BL21 | In this study |
| His-sFap1-GlcNAc-Glc(-Glc)-GlcNAc | pET-28b: sFap1 and pvpt-Gtf123-T1 co-transformed into BL21 | In this study |
| GST-DUF1792 | pGEx-5x-1: DUF1792 transformed into Top10 | 27 |
| GST-dGT1 | pGEx-5x-1: dGT1 transformed into Top10 | 27 |
| GST-CgT | pGEx-5x-1: CgT transformed into Top10 | In this study |
| Plasmids | ||
| pvpt-hsv-his | E. coli-streptococci shuttle vector; Ermr | 45 |
| pET-sumo | His-SUMO fusion protein expression vector; Kanr | 25 |
| pET-28b | His fusion protein expression vector; Kanr | Amersham |
| pGEx-6p-1 | GST fusion protein expression vector; Ampr | Amersham |
| pET-sumo: CgT | CgT cloned in pET-sumo; Kanr | In this study |
| pET-28b: CgT | CgT cloned in pET-28b; Kanr | In this study |
| pET-sumo: CgTΔH1–3 | Deletion of whole helix domain from pET-sumo: CgT; Kanr | In this study |
| pET-sumo: CgT-DH1 | Deletion of first helix domain from pET-sumo: CgT; Kanr | In this study |
| pET-sumo: CgT-DH2 | Deletion of second helix domain from pET-sumo: CgT; Kanr | In this study |
| pET-sumo: CgT-DH3 | Deletion of third helix domain from pET-sumo: CgT; Kanr | In this study |
| pvpt-dGT1 | dGT1 cloned into pvpt-hsv-his; Ermr | In this study |
| pvpt-DUF1792 | DUF1792 cloned into pvpt-hsv-his; Ermr | In this study |
| pvpt-CgT | CgT cloned into pvpt-hsv-his; Ermr | In this study |
| pvpt-DUF1792-pmal-CgT | CgT with promoter cloned into pvpt-DUF1792; Ermr | In this study |
| pvpt-Gtf123 | Gtf12 and Gtf3 cloned into pvpt-hsv-his; Ermr | 27 |
| pvpt-Gtf123-DUF1792 | Gtf12 and Gtf3 and DUF1792 cloned into pvpt-hsv-his; Ermr | In this study |
| pvpt-Gtf123-T1 | Gtf12 and Gtf3 and dGT1 cloned into pvpt-hsv-his; Ermr | In this study |
| pvpt-dGT-AxD | Site-direct mutant Asp378 to Ala from pvpt-dGT1; Ermr | In this study |
| pvpt-dGT-DxA | Site-direct mutant Asp380 to Ala from pvpt-dGT1; Ermr | In this study |
| pGEx-6p-1: DUF1972 | DUF1972 cloned into pEGx-6p-1; Ampr | 27 |
| pGEx-6p-1: dGT1 | dGT1 cloned into pEGx-6p-1; Ampr | 27 |
| pGEx-6p-1: CgT | CgT cloned into pEGx-6p-1; Ampr | In this study |