Table 1.
Bacterial strains and plasmids used in this study
| Bacterial strain or plasmid | Description or relevant genotype or phenotype | Reference or source |
|---|---|---|
| A. actinomycetemcomitansstrains | ||
| VT1169 | Wild-type strain, a derivative of SUNY465 | 31 |
| KM73 | emaA mutant strain | 28 |
| VT1565 | aae mutant strain | 41 |
| KM333 | In-frame deletion of emaA gene region corresponding to deletion of amino acids 16 to 39 of EmaA protein | This study |
| E. coli strains | ||
| DH10B | F−mcrA Δ(mrr-hsd RMS-mcr BC) φ80dlacZΔM15 Δlac X74 end A1 rec A1 deo R Δ(ara leu)7697 ara D139 gal U gal K nup G rps L λ− | 15 |
| DH5α λ pir | endA1 hsdR17(r− m+) supE44 thi-1 recA1 gyrA1(Nalr) relA1Δ(lacIZYA-argF)U169 deo[φ80 ΔlacD(lacZ)M15] pir R6K | |
| SM10 λ pir | thi-1 thr leu tonA lacY supE recA::RP4-2Tc::Mu l pir R6K | 39 |
| CC118 | phoA20 thi-1 rspE rpoB argE(Am) recA1 | 18 |
| Plasmids | ||
| pKM1 | Shuttle vector; Kanr | 42 |
| pKM2 | Shuttle vector; Cmr | 14 |
| pVT1460 | Mobilization plasmid | 29 |
| pVT1566 | aae gene sequence cloned into pGEM-T Easy | 41 |
| pKM9 | Upstream promoter region and emaA in pKM1 | 42 |
| pQE30 | Intermediate cloning vector; Ampr | Qiagen |
| pHRM104 | Plasmid containing truncated phoA | 17 |
| pQEmaA | Upstream promoter region and emaA in pQE30 | This study |
| pKMΔ2-23 | In-frame deletion of bp 4 to 69 of emaA in pKM9 | This study |
| pKMΔ2-35 | In-frame deletion of bp 4 to 105 of emaA in pKM9 | This study |
| pKMΔ24-53 | In-frame deletion of bp 70 to 159 of emaA in pKM9 | This study |
| pKMΔ16-39 | In-frame deletion of bp 46 to 117 of emaA in pKM9 | This study |
| pKMOmp34SP | emaA signal peptide replaced by omp34 signal peptide in pKM9 | This study |
| pKMEmaASP-Aae | aae containing the emaA signal peptide and upstream promoter region in pKM2 | This study |
| pHRMEmaASP | emaA signal peptide and upstream promoter region in pHRM104 | This study |
| pHRMEmaASPΔ2-56 | In-frame deletion of bp 4 to 168 of pHRMEmaASP | This study |
| pHRMEmaASPΔ2-23 | In-frame deletion of bp 4 to 69 of pHRMEmaASP | This study |
| pHRMEmaASPΔ2-35 | In-frame deletion of bp 4 to 105 of pHRMEmaASP | This study |
| pHRMEmaASPΔ24-53 | In-frame deletion of bp 70 to 159 of pHRMEmaASP | This study |
| pHRMEmaASPΔ16-39 | In-frame deletion of bp 46 to 117 of pHRMEmaASP | This study |
| pHRMHiaSP | hia signal peptide and emaA upstream promoter region in pHRM104 | This study |