TABLE 3.
Bacterial strains and plasmids used in this research
| Species and strain or plasmid | Feature(s) | Reference or source |
|---|---|---|
| Escherichia coli | ||
| DH5α | Cloning strain | 73 |
| K-12 MG1655 | Model E. coli strain | 74 |
| BL21(DE3) pLys | Engineered E. coli strain for protein expression, contains Tn10 that produces T7 polymerase and plasmid pLys; the presence of pLys is maintained with 5 μg/ml chloramphenicol | Novagen |
| Streptococcus mitis | ||
| ATCC 49456 (SM61) | Type strain of S. mitis | ATCC |
| SM61 ΔcdsA | SM61 with coding region of cdsA (SM12261_RS08390) deleted | This study |
| SM61 ΔltaS | SM61 with coding region of ltaS (SM12261_RS03435) replaced with gene ermB | This study |
| Streptococcus sp. | ||
| 1643 (SM43) | Mitis group Streptococcus isolated from infective endocarditis patient | 42 |
| SM43 ΔcdsA | SM43 with coding region of cdsA (FD735_RS08600) deleted | 18 |
| SM43 ΔpgsA | SM43 with coding region of pgsA (FD735_RS09695) replaced with gene ermB | 18 |
| Streptococcus oralis | ||
| ATCC 35037 | Type strain of S. oralis | ATCC |
| 1647 | Isolated from infective endocarditis patient | 42 |
| Streptococcus pneumoniae | ||
| D39 | Clinically isolated strain, serotype 2 | 75 |
| TIGR4 | Clinically isolated strain, serotype 4 | 76 |
| Streptococcus pyogenes | ||
| NZ131 | Clinically isolated strain, serotype M49 | ATCC |
| Streptococcus agalactiae | ||
| A909 | Isolated from a septic human neonate, serotype Ia | ATCC |
| Staphylococcus aureus | ||
| ANG113 | Strain RN4220, isogenic wild-type control of ANG499 | 36 |
| ANG499 | Generated from strain RN4220 (wild type), expression of chromosomal ltaS is induced with 1 mM IPTG; the genotype maintained with 5 μg/ml erythromycin | 36 |
| Enterococcus faecalis | ||
| OG1RF | Rifampin- and fusidic acid-resistant derivative of a human oral cavity isolate | 77 |
| Plasmids | ||
| pABG5 | Low-copy-number shuttle plasmid; confers kanamycin resistance | 68 |
| pitetR-ltaS | pABG5 with S. mitis ltaS coding region under control of tetracycline-inducible promoter Pxyl/tet | This study |
| pitetR-SAltaS | pABG5 with S. aureus ltaS (SAV0719) coding region under control of promoter Pxyl/tet | This study |
| pitetR | pitetR-ltaS that lacks the ltaS coding region and has EcoRI introduced; serves as empty plasmid control | This study |
| pET-28a(+) | Expression plasmid; confers kanamycin resistance | Novagen |
| pET-ltaS | pET-28a(+) with S. mitis ltaS under control of IPTG-inducible promoter | This study |
| pMSP3535 | Confers erythromycin resistance; used to obtain ermB gene | 7 |