TABLE 3.
Strains and plasmids used in the study
| Strain, vector, or plasmid | Description | Reference(s) or source |
|---|---|---|
| Strains | ||
| USA300 LAC* | S. aureus USA300 LAC lacking pUSA03 (designated as the wild type [WT]) | 22, 51 |
| RN4220 | S. aureus restriction deficient, cloning intermediate | 56 |
| ΦΝΞ07490 | S. aureus Newman with Bursa aurealis insertion at nucleotide 602 of essH | 37 |
| essH | S. aureus USA300 LAC* essH::ermB | This study |
| essB | S. aureus USA300 LAC* ΔessB | 24 |
| sagB | S. aureus USA300 LAC* ΔsagB::aphA | 32 |
| E. coli DH5α | F− ϕ80lacZΔM15 Δ(lacZYA-argF)U169 recA1 endA1 hsdR17(rK− mK+) pho A supE44 λ− thi-1 gyrA96 relA1 | Our collection |
| E. coli BL21 | B F− ompT gal dcm lon hsdSB(rB−mB−) [malB+]K-12(λS) | 57 |
| Vectors and plasmids | ||
| pOS1 | E. coli/S. aureus shuttle vector | 58 |
| pSEW016 | pWWW412 with a modified polylinker herein referred as vector | This study; 52 |
| pET15b | E. coli vector for production of recombinant proteins | Addgene |
| pessH | essH open reading frame cloned into pSEW016 | This study |
| pessHC199A | essHC199A cloned into pSEW016 | This study |
| pessHH254A | essHH254A cloned into pSEW016 | This study |
| pessHC199A/H254A | essHC199A/H254A cloned into pSEW016 | This study |
| pessHSe | S. epidermidis 14.1.R1 essH cloned into pSEW016 | This study |
| pessHSe(SP) | S. aureus USA300 LAC essH nucleotides 1 to 78 fused to S. epidermidis 14.1.R1, essH nucleotides 85 to 960 cloned into pSEW016 | This study |
| pSTREPessH | S. aureus USA300 LAC essH with STREP II tag sequence inserted between nucleotides 72 and 73 and cloned into pSEW016 | This study |
| psagB | sagB cloned into pOS1 | 32 |
| pET15b-STREPessH | essH nucleotides 73 to 894 with N-terminal STREP II tag cloned into pET15b | This study |
| pET15b-STREPessHC199A/H254A | essHC199A/H254A nucleotides 73 to 894 with N-terminal STREP II cloned into pET15b | This study |