TABLE 1.
Strains and plasmids used in the study
| Relevant properties | Source/reference | |
|---|---|---|
| Strains | ||
| S. parasanguinis FW213 | Wild type | |
| E. coli Top10 | Host for propagation of recombinant plasmids | Invitrogen |
| E. coli BL21 Gold (DE3) | pET system host strain | Invitrogen |
| fap1 mutant | Wild type; fap1::aphA3; Kanr | Ref. 5 |
| gtf1 mutant | VT508 | Ref. 64 |
| gtf3 mutant | Wild type; gtf3 knockout; gtf3::aphA3; Kanr | Ref. 17 |
| dGT1 mutant | Wild type; dGT1 knockout; dGT1::aphA3; Kanr | Ref. 15 |
| galT2 mutant | Wild type; galT2 knockout; galT2::aphA3; Kanr | Ref. 22 |
| gly mutant | Wild type; gly knockout; gly::aphA3; Kanr | This study |
| His-Gly | pET28b-Gly in BL21 | This study |
| rFap1 | pET28a-rFap1-R1 in BL21 | Ref. 15 |
| rFap1-Gtf123-dGT1-Gly | pET28a-rFap1-R1 and pvpt-Gtf123-dGT1-Gly and pHSG576 co-transformed into BL21 | This study |
| rFap1-Gtf123-dGT1-GalT2 | pET28a-rFap1-R1 and pvpt-Gtf123-dGT1-GalT2 and pHSG576 co-transformed into BL21 | This study |
| rFap1-Gtf123-dGT1-GalT2-Gly | pET28a-rFap1-R1 and pvpt-Gtf123-dGT1-GalT2 and pHSG576-Gly co-transformed into BL21 | Ref. 15 |
| Plasmids | ||
| pET28a | His tag fusion protein expression vector Kanr | Invitrogen |
| pHSG576 | PCR cloning vector; Cmr | Ref. 5 |
| pALH124 | aphA3 Kanr cassette-containing plasmid; Ampr Kanr | Ref. 65 |
| pVPT-CHSV | E. coli-Streptococcus shuttle vector; Ermr | Ref. 66 |
| pVPT-Gtf123-dGT1-Gly | Gtf123, dGT1 and Gly cloned into pVPT-CHSV; Ermr | This study |
| pET28b-Gly | Gly cloned into pET28b | This study |
| pVPT-Gly | Gly cloned into pVPT-CHSV; Ermr | This study |