Table 1.
Bacterial strains and plasmids used in this study
| Strain | Plasmid | Function | Restriction sites used for cloning | Primer used for PCR amplification or site-directed mutagenesisa |
|---|---|---|---|---|
| SA113 | pMADΔyjbH | Allelic replacement of yjbH | SacI/BamHI | 5′GAAGCAATCGGAGCTCAATGGATCGA |
| Upstream fragment | 5′CGATACTACGGATCCTGAGTTAGAGTTCG | |||
| NcoI/XbaI | 5′AATTCATGCCATGGAAGAAGCAGGTG | |||
| Downstream fragment | 5′CCAGATGGCGATTTCTAGAAATCTAAAATGCC | |||
| SA113ΔyjbH | pRB474yjbH | Complementation of mutant strain with wild-type copy of S. aureus yjbH | PstI/XbaI | 5′GAGCGCTTAAGATTAACTGCAGATCATATGGTG |
| 5′CTCGTTACATCTAGAGTGTACGAGGTC | ||||
| SA113ΔyjbH | pRB474yjbHBs | Complementation of mutant strain with wild-type copy of B. subtilisyjbH | HindIII/BamHI | 5′ATCAAGCTTAAAGGAGGTATCATCATGACAAACTATCAGCATGAGCTATAC |
| 5′CATATGGATCCCTGCGGCTATTTTTCACATG | ||||
| SA113 | pRB474 | Control wild-type strain with empty vector | ||
| SA113ΔyjbH | pRB474 | Control mutant strain with empty vector | ||
| SA113ΔyjbH | pRB474yjbHC2 (C114G, C116G) | Complementation of mutant strain with mutated yjbH | 5′CAATACCTGCATTTTGAATACCGTCACCAATCATTGATTCTG | |
| 5′CAGAATCAATGATTGGTGACGGTATTCAAAATGCAGGTATTG | ||||
| SA113ΔyjbH | pRB474yjbHC3a (C30G, C114G, C116G) | Complementation of mutant strain with mutated yjbH | 5′GATTGCTGATAATTTGAAGCCATCGGAGCTAAATGGATC | |
| 5′GATCCATTTAGCTCCGATGGCTTCAAATTATCAGCAATC | ||||
| SA113ΔyjbH | pRB474yjbHC3b (C64G, C114G, C116G) | Complementation of mutant strain with mutated yjbH | 5′GGATGTACTTTGAGCTTGGCCTTTCGTTAATACTTTTAACG | |
| 5′CGTTAAAAGTATTAACGAAAGGCCAAGCTCAAAGTACATCC | ||||
| SA113ΔyjbH | pRB474yjbHC4 (C30G, C64G, C114G, C116G) | Complementation of mutant strain with mutated yjbH | 5′GATTGCTGATAATTTGAAGCCATCGGAGCTAAATGGATC | |
| 5′GATCCATTTAGCTCCGATGGCTTCAAATTATCAGCAATC |
a
Restriction sites in primers used for cloning or mutated nucleotides in primers used for site-directed mutagenesis are indicated in boldface.