Table 1.
Strains and plasmids.
| Strain or plasmid | Description | Source or reference |
|---|---|---|
| B. mallei | ||
| ATCC 23344 | Wild-type strain; polymyxin B resistant, kanamycin and zeocin sensitive | [10] |
| pal KO | Isogenic pal mutant strain of ATCC 23344; resistant to polymyxin B and zeocin, sensitive to kanamycin | This study |
| E. coli | ||
| EPI300 | Strain used for general recombinant DNA manipulations; sensitive to kanamycin, zeocin, tetracycline, and chloramphenicol | epicenter/Lucigen |
| EC100D pir+ | Strain used for recombinant DNA manipulations of plasmid pKAS46 and derivatives; sensitive to kanamycin and zeocin | epicenter/Lucigen |
| S17 | Strain used for conjugational transfer of plasmids to B. mallei; sensitive to kanamycin, zeocin, and polymyxin B | [34] |
| TUNER | Protein expression strain used to purify His-tagged PalBm and LolCBm proteins; chloramphenicol sensitive | Millipore Sigma |
| Plasmids | ||
| pBHR1 | Cloning vector; confers resistance to chloramphenicol and kanamycin | MoBiTec |
| pBHR1∆ Dra | pBHR1 containing a 339 nt deletion in the chloramphenicol resistance marker; confers resistance only to kanamycin | [43] |
| pPalBm | pBHR1 in which a 812 nt DNA fragment containing the pal gene of B. mallei ATCC 23344 was inserted; confers resistance only to kanamycin | This study |
| pEM7/ZEO | Source of the zeocin resistance cassette; confers resistance to zeocin | ThermoFisher Scientific |
| pACYC184 | Cloning vector; confers resistance to chloramphenicol and tetracycline | New England Biolabs Inc. |
| pACYCPalBm | pACYC184 in which a 1,332 nt DNA fragment containing the pal gene of B. mallei ATCC 23344 was inserted; confers resistance to tetracycline | This study |
| pACYCPalBmZEO | pACYCPalBm in which a zeocin resistance cassette was inserted near the middle of the pal gene; confers resistance to tetracycline and zeocin | This study |
| pKAS46 | Gene replacement vector; confers resistance to kanamycin | [35] |
| pKASPalBmZEO | pKAS46 containing the pal gene interrupted with a zeocin resistance cassette from pACYCPalBmZEO; confers resistance to kanamycin and zeocin | This study |
| pETcoco-1 | His-tagged protein expression vector; confers resistance to chloramphenicol | Millipore Sigma |
| pHisPalBm | pETcoco-1 in which a gene fragment encoding amino acids 22 to 170 of B. mallei ATCC 23344 Pal was inserted; confers resistance to chloramphenicol | This study |
| pHisLolCBm | pETcoco-1 in which a gene fragment encoding amino acids 47 to 243 of B. mallei ATCC 23344 LolC was inserted; confers resistance to chloramphenicol | This study |
| pMHD11 | pZL185 (PIV5 genome) in which a gene fragment encoding amino acids 22 to 170 of B. mallei ATCC 23344 Pal was inserted; confers resistance to chloramphenicol | This study |
| Viruses | ||
| PIV5-PalBm (MHD11) | PIV5 expressing amino acids 22 to 170 of B. mallei ATCC 23344 Pal protein | This study |
| PIV5-BatA | PIV5 expressing amino acids 30 to 307 of B. mallei ATCC 23344 BatA protein | [41] |
| PIV5-Tb | PIV5 expressing Ag85B protein of M. tuberculosis | [40] |