Table 1.
Bacterial strains and plasmids used in this study
| Strain/plasmid | Revelant genotype | Characteristics | Source/reference |
|---|---|---|---|
| Strains | |||
| RN4220 | 8325-4; r− m+; restriction-defective laboratory strain | 72 | |
| ISP794 | 8325 pig 131; rsbU mutant | 73 | |
| AR852 | ISP794; rsbU+ | ISP794; (rsbUVW-sigB)+ tetL nearby | 24 |
| AR376 | ISP4-2-1 | GISA in vitro-derived strain | 14 |
| AR774 | vraS (G45R) | ISP794; vraS (G45R); Kanr nearby | 10 |
| AR826 | stp1 (Q12stop) | ISP794, stp1 (Q12stop); Kanr nearby | 10 |
| AR1079 | yjbH (K23stop) | ISP794, yjbH (K23stop); Eryr nearby | 10 |
| Δspx | 8325-4-derived strain | spx-deleted strain | 30 |
| spx+ | 8325-4 Δspx Pspx-spx::geh | spx-deleted strain chromosomally complemented with the intact copy of spx inserted into the geh locus | 30 |
| Δspx rsbU+ | 8325-4-derived strain | spx-deleted strain; (rsbUVW-sigB)+; tetL nearby | This study |
| AR612 | ISP4-2-1; ΔtrfA | ISP4-2-1; trfA::tetK | 14 |
| AR916 | ISPΔvraSR | ISP794; ΔvraSR::Kanr | 74 |
| Pair 1 non-GISA | Clinical MRSA glycopeptide-susceptible strain of pair 1 | 27 | |
| Pair 1 GISA | Clinical MRSA glycopeptide-resistant strain of pair 1 | 27 | |
| Pair 2 non-GISA | Clinical MRSA glycopeptide-susceptible strain of pair 2 | 27 | |
| Pair 2 GISA | Clinical MRSA glycopeptide-resistant strain of pair 2 | 27 | |
| Pair 3 non-GISA | Clinical MRSA glycopeptide-susceptible strain of pair 3 | 27 | |
| Pair 3 GISA | Clinical MRSA glycopeptide-resistant strain of pair 3 | 27 | |
| Pair 4 non-GISA | Clinical MRSA glycopeptide-susceptible strain of pair 4 | 27 | |
| Pair 4 GISA | Clinical MRSA glycopeptide-resistant strain of pair 4 | 27 | |
| Pair 5 non-GISA | Clinical MRSA glycopeptide-susceptible strain of pair 5 | 27 | |
| Pair 5 GISA | Clinical MRSA glycopeptide-resistant strain of pair 5 | 27 | |
| BL2 α(DE3) | E. coli | IPTG-inducible T7 RNA polymerase | New England Biolabs |
| Plasmids | |||
| pBluescript II KS(+) | Cloning vector; Ampr | ||
| pAM845 | pKS+ containing a Kpn-Pst fragment coding for TrfA | This study | |
| pTYB12 | N-terminal fusion IMPACT intein and chitin binding domain plasmid | New England Biolabs | |
| pAM873 | pTYB12 containing trfA cloned into NdeI-PstI restriction sites | This study |