TABLE 1.
Strains and plasmids used in this study
| Plasmid or strain | Relevant characteristic(s)a | Source or reference |
|---|---|---|
| Strains | ||
| E. coli | ||
| TOP10 | Invitrogen | |
| SCS110 | Stratagene | |
| B. anthracis | ||
| Ames 33 | pXO1− pXO2− | 32 |
| Ames 35 | pXO1+ pXO2− | 32 |
| A33(pSW4-GFPmut1) | Ames 33 electroporated with pSW4-GFPmut1, Kmr | This study |
| A33(pSW4-GFPopt) | Ames 33 electroporated with pSW4-GFPopt, Kmr | This study |
| A33(pSW4-CFP) | Ames 33 electroporated with pSW4-CFP, Kmr | This study |
| A33(pSW4-CFPopt) | Ames 33 electroporated with pSW4-CFPopt, Kmr | This study |
| A33(pSW-PAGFPopt) | Ames 33 electroporated with pSW4-PAGFPopt, Kmr | This study |
| S. aureus | ||
| RN4220 | Hemolysin-defective S. aureus strain, Spr | 13 |
| RN(pTetONGFPopt) | RN4220 electroporated with pTetONGFPopt, Spr | This study |
| RN(pTetONCFPopt) | RN4220 electroporated with pTetONCFPopt, Spr | This study |
| RN(pTetONYFPopt) | RN4220 electroporated with pTetONYFPopt, Spr | This study |
| Plasmids | ||
| pCR2.1 TOPO | Cloning vector, Kmr Apr | Invitrogen |
| pUC18-GFPopt | Vector harboring custom codon-optimized GFPmut1, Apr | BlueHeron |
| pS10-CFP | pCR2.1 harboring CFP under the control of S10 promoter of S. aureus | This study |
| pS10-CFPopt | pCR2.1 harboring CFPopt under the control of S10 promoter of S. aureus | This study |
| pSW4 | Gram-positive/gram-negative shuttle vector with PA promoter, Kmr Apr | 32 |
| pSW4-GFPmut1 | pSW4 plasmid containing 729-bp AseI/BamHI-cloned gfpmut1 | This study |
| pSW4-GFPopt | pSW4 plasmid containing 729-bp AseI/BamHI-cloned gfpopt | This study |
| pSW4-CFP | pSW4 plasmid containing 732-bp AseI/BamHI-cloned cfp | This study |
| pSW4-CFPopt | pSW4 plasmid containing 732-bp AseI/BamHI-cloned cfpopt | This study |
| pSW4-YFP | pSW4 plasmid containing 732-bp AseI/BamHI-cloned yfp | This study |
| pSW4-YFPopt | pSW4 plasmid containing 729-bp AseI/BamHI-cloned yfpopt | This study |
| pSW4-PAGFP | pSW4 plasmid containing 732-bp AseI/BamHI-cloned pagfp | This study |
| pYJ335 | Vector with tetracycline-inducible Pxyl/tetO promoter | 20 |
| pJRS312 | Vector containing spectinomycin cassette | 36 |
| pTetON | pYJ335 derivative with inactive tetracycline repressor promoter, Apr Spr | This study |
| pTetONGFPopt | pTetON containing 729-bp gfpopt cloned via SmaI/SbfI | This study |
| pTetONCFPopt | pTetON containing 729-bp cfpopt cloned via SmaI/SbfI | This study |
| pTetONYFPopt | pTetON containing 729-bp cfpopt cloned via SmaI/SbfI | This study |
a
Abbreviations: Apr, ampicillin resistance; Kmr, kanamycin resistance; Spr, spectinomycin resistance.