Table 2.
Bacterial strains used in this study
| Strain | Relevant features | Reference |
|---|---|---|
| Escherichia coli strains | ||
| XL1 Blue | Cloning strain, TetR – ANG127 | Stratagene |
| DH5α | Cloning strain – ANG397 | Hanahan (1983) |
| Rosetta | Strain used for protein expression – ANG574 | Novagen |
| ANG201 | pCN34 in E. coli; source for Kan (aphA-3) marker; KanR and AmpR | Charpentier et al. (2004) |
| ANG203 | pCN49 in E. coli; source for Erm (ermC) marker; AmpR | Charpentier et al. (2004) |
| ANG204 | pCN55 in E. coli; source for Spec (add9) marker; AmpR | Charpentier et al. (2004) |
| ANG243 | pCL55 in XL1 Blue; S. aureus single-site integration vector; AmpR | Lee et al. (1991) |
| ANG284 | pitet in XL1 Blue; E. coli / S. aureus shuttle vector with tetracycline inducible promoter; AmpR | Gründling and Schneewind (2007b) |
| ANG503 | pCL55-ltaS in XL1 Blue; AmpR | This study |
| ANG506 | pCL55-yqgS in XL1 Blue; AmpR | This study |
| ANG508 | pitet-ltaS in XL1 Blue; ltaS under tetracycline inducible promoter; AmpR | Gründling and Schneewind (2007a) |
| ANG509 | pitet-yfnI in XL1 Blue; yfnI under tetracycline inducible promoter; AmpR | Gründling and Schneewind (2007a) |
| ANG510 | pitet-yflE in XL1 Blue; yflE under tetracycline inducible promoter; AmpR | Gründling and Schneewind (2007a) |
| ANG512 | pitet-yvgJ in XL1 Blue; yvgJ under tetracycline inducible promoter; AmpR | Gründling and Schneewind (2007a) |
| ANG1444 | pProEX-eYflE in DH5α; plasmid for overexpression of eYflE (eLtaSBS) domain; N-terminal His-tag; AmpR | This study |
| ANG1445 | pProEX-eYfnI in DH5α; plasmid for overexpression of eYflnI domain; N-terminal His-tag; AmpR | This study |
| ANG1446 | pProEX-eYvgJ in DH5α; plasmid for overexpression of eYvgJ domain; N-terminal His-tag; AmpR | This study |
| ANG1447 | pProEX-eYqgS in DH5α; plasmid for overexpression of eYqgS domain; N-terminal His-tag; AmpR | This study |
| ANG1474 | pProEX-eYflE in Rosetta strain; strain use for overexpression of eYflE (eLtaSBS) protein with N-terminal His tag; AmpR | This study |
| ANG1475 | pProEX-eYfnI in Rosetta strain; strain use for overexpression of eYfnI protein with N-terminal His tag; AmpR | This study |
| ANG1476 | pProEX-eYvgJ in Rosetta strain; strain use for overexpression of eYvgJ protein with N-terminal His tag; AmpR | This study |
| ANG1477 | pProEX-eYqgS in Rosetta strain; strain use for overexpression of eYqgS protein with N-terminal His tag; AmpR | This study |
| ANG1512 | pCN34itet-ltaS in XL1 Blue; ltaS under tetracycline inducible promoter; KanR, AmpR | This study |
| ANG1514 | pCN34itet-yfnI in XL1 Blue; yfnI under tetracycline inducible promoter; KanR, AmpR | This study |
| ANG1615 | pitet-RBltaS-yqgS in XL1 Blue; yqgS with ltaS ribosomal binding site under tetracycline inducible promoter; AmpR | This study |
| ANG1652 | pCN34itet-yqgS in XL1 Blue; yqgS under tetracycline inducible promoter; KanR, AmpR | This study |
| ANG1656 | pCN34itet-yvgJ in XL1 Blue; yvgJ under tetracycline inducible promoter; KanR, AmpR | This study |
| ANG1660 | pCN34itet-yflE in XL1 Blue; yflE (ltaSBS) under tetracycline inducible promoter; KanR, AmpR | This study |
| ANG1676 | pCN38 in E. coli; source for Gram-positive Cam (cat194) marker; AmpR | Charpentier et al. (2004) |
| Staphylococcus aureus strains | ||
| RN4220 | Transformable laboratory strain | Kreiswirth et al. (1983) |
| ANG499 | RN4220 with IPTG-inducible ltaS expression; ErmR, IPTG | Gründling and Schneewind (2007a) |
| ANG514 | pitet-ltaS integrated in strain ANG499; ErmR, CamR, IPTG | Gründling and Schneewind (2007a) |
| ANG1130 | ANG499 with pCN34; ErmR, KanR, IPTG | This study |
| ANG1571 | ANG499 with pCN34itet-ltaS; ErmR, KanR, IPTG | This study |
| ANG1573 | ANG499 with pCN34itet-yfnI; ErmR, KanR, IPTG | This study |
| ANG1654 | ANG499 with pCN34itet-yqgS; ErmR, KanR, IPTG | This study |
| ANG1658 | ANG499 with pCN34itet-yvgJ; ErmR, KanR, IPTG | This study |
| ANG1662 | ANG499 with pCN34itet-yflE; ErmR, KanR, IPTG | This study |
| Bacillus subtilis strains | ||
| B. subtilis | Bacillus subtilis 168 – Transformable lab strain, trpC2 – ANG1691 | Burkholder and Giles (1947) |
| ANG1692 | Bacillus subtilis 168 yfnI::Cam (ΔyfnI) | This study |
| ANG1693 | Bacillus subtilis 168 yflE::Kan (ΔltaSBS) | This study |
| ANG1694 | Bacillus subtilis 168 yqgS::Spec (ΔyqgS) | This study |
| ANG1695 | Bacillus subtilis 168 yvgJ::Erm (ΔyvgJ) | This study |
| ANG1696 | Bacillus subtilis 168 yflE::Kan, yqgS::Spec, yvgJ::Erm (yfnI only) | This study |
| ANG1697 | Bacillus subtilis 168 yfnI::Cam, yqgS::Spec, yvgJ::Erm (ltaSBS only) | This study |
| ANG1698 | Bacillus subtilis 168 yfnI::Cam, yflE::Kan, yvgJ::Erm (yqgS only) | This study |
| ANG1699 | Bacillus subtilis 168 yfnI::Cam, yflE::Kan, yqgS::Spec (yvgJ only) | This study |
| ANG1701 | Bacillus subtilis 168 yfnI::Cam, yflE::Kan, yqgS::Spec, yvgJ::Erm (4xΔ) | This study |
| ANG1702 | Bacillus subtilis 168 yflE::Kan, yfnI::Cam (express-yvgJ/yqgS) | This study |
| ANG1703 | Bacillus subtilis 168 yflE::Kan, yqgS::Spec (express-yvgJ/yfnI) | This study |
| ANG1704 | Bacillus subtilis 168 yfnI::Cam, yqgS::Spec (express-yvgJ/yflE) | This study |
| L5703 | Bacillus subtilis with ribitol-Pi wall teichoic acid (ribitol-Pi WTA) | Karamata et al. (1987) |
Antibiotics were used at the following concentrations: for E. coli cultures: ampicillin (AmpR) 100 µg ml−1; kanamycin (KanR) 30 µg ml−1; tetracycline (TetR) 10 µg ml−1; for S. aureus cultures: erythromycin (ErmR) 10 µg ml−1; chloramphenicol (CamR) 7.5 to 10 µg ml−1 and IPTG at 1 mM; for B. subtilis cultures: erythromycin 5 µg ml−1; chloramphenicol 10 µg ml−1; kanamycin 10 µg ml−1; spectinomycin (Spec) 100 or 200 µg ml−1.