Table 1.
Strains, plasmids, and primers
| Description | Source | |
|---|---|---|
| Strains | ||
| AH1263 | S. aureus strain USA300–0114, Erm-sensitive | 46 |
| JLB31 | fakB1::ΦΝΣ ΔfakB2 of strain AH1263 | 1 |
| Sa178RI | Derived from S. aureus strain RN4220 | 47 |
| TIGR4 | Wildtype encapsulated S. pneumoniae | www.tigr.org |
| JMG1 | TIGR4 with fakB3 replaced with Kan cassette | This study |
| Plasmids | ||
| pG164 | Isopropyl 1-thio-β-d-galactopyranoside-inducible vector | 47 |
| pCS119 | pCM28SarAP1promoter | 48 |
| pPJ480 | pCS119 vector with RBS site | This study |
| pET28a | Expression vector | Novagen |
| pET28SpA | His tag SpFakA expression vector | This study |
| pET28SpB1 | His tag SpFakB1 expression vector | This study |
| pET28SpB2 | His tag SpFakB2 expression vector | This study |
| pET28SpB3 | His tag SpFakB3 expression vector | This study |
| pSaB1 | pCS119 expressing SaFakB1 | This study |
| pSaB2 | pCS119 expressing SaFakB2 | This study |
| pSpB1 | pPJ480 expressing SpFakB1 | This study |
| pSpB2 | pPJ480 expressing SpFakB2 | This study |
| pSpB3 | pPJ480 expressing SpFakB3 | This study |
| pDL278 | E. coli, Gram-positive shuttle vector | 49 |
| pSpFakB3 | SpFakB3 S. pneumoniae expression plasmid | This study |
| pABG5 | E. coli, Gram-positive shuttle vector | 34 |
| Primers | Sequence (5′ to 3′) | |
| Primer 1 | CTCAAGTGGTAAATGGGTCAA | |
| Primer 2 | AAATGGTTCGCTGGGTTTATCAGAAATTCTCCATCTTTGTCA | |
| Primer 3 | GATAAACCCAGCGAACCATTT | |
| Primer 4 | ATACAAATTCCTCGTAGGCGC | |
| Primer 5 | GCGCCTACGAGGAATTTGTATTTGCATTCTTGACAAGAGGTG | |
| Primer 6 | ATCATAATGCAGACTTCCCGC | |
| Primer 7 | TGGAGAATTTCTATGACTTGG | |
| Primer 8 | ATGCAATTAATCAATTTC | |
| pABGB3BamH1 | CTTGGATCCATTTTCTCTCCTCTCAAA | |
| pABGB3Sac1 | GATGAGCTCTTAATCAATTTCATAGCC |