TABLE 1.
Plasmid competitors used as internal standards for RT-PCR and related materials
| Plasmida | Size (kb) | Properties and construction methodb | Reference |
|---|---|---|---|
| pCU1 | 4.9 | Shuttle vector for E. coli and S. aureus | 1 |
| pCU hlg | 10.3 | pCU1; 5.4-kb ScaI DNA fragment containing hlgA, -B, and -C genes in HincII-PstI sites of pCU1 plasmid | 31 |
| pCU hlgA | 6.9 | pCU1; 2-kb EcoRI-EcoRI DNA fragment encoding hlgA | This study |
| pCU lukED | 8.1 | pCU1; 3.2-kb HindIII DNA fragment encoding lukE and lukD | 16 |
| pCU lukPV | 8.0 | pCU lukS-PV/lukFPV; HindIII-HincII sites containing 3.1-kb HindIII-NruI fragment containing lukS-PV and lukFPV | 31 |
| pGP7 | 7.0 | pCR II lukPV (NdeI°, 80 nt); 80-bp MseI-MseI fragment from pUC19 inserted into NdeI site of pCR II-lukPV (encoding structural lukS-PV and lukFPV genes obtained with TA cloning kit [In vitrogen Corporation]) | This study |
| pGP8 | 10.2 | pCU hlgACB (deletion of nt 2487 to 2598); deletion of pCU-hlgACB at positions 2487 to 2598 (hlgC) obtained by Pfu DNA polymerase (Stratagene) amplification with dedicated primers surrounding the deletion | This study |
| pGP9 | 7.1 | pCU hlgA (EcoRV°, 200 nt); 200-bp AluI-AluI fragment from pCU1 inserted at the unique EcoRV site at position 275 of hlgA | This study |
| pGP10 | 8.35 | pCU lukED (EcoRI°, 250 nt); 250-bp Tsp509I-Tsp509I from pCU1 hlg inserted at the unique EcoRI site at position 1855 of lukD | This study |
a
Presence of plasmids was maintained in cultures by using 100 μg of ampicillin/ml of medium.
b
nt, nucleotide.