Table 1 List of strains and plasmids used in this study
Strains/Plasmids | Description | Source |
|---|---|---|
K. pneumoniae strains | ||
MGH 78578 | Clinical isolate from a pneumonia patient. Parental (wild-type) strain for gene deletion. | ATCC |
∆wzabc | In-frame deletion of wzabc operon (KPN_02510-KPN_02512) | This study |
∆cps | In-frame deletion of capsule biosynthesis cluster ranging from ugd (KPN_02493) to galF (KPN_02515) | This study |
E. coli strains | ||
Top10 | Competent cells for general cloning | Life Technologies |
Plasmids | ||
pACBSR | A p15A replicon plasmid containing an arabinose-inducible λ-Red recombinase and chloramphenicol resistance selection marker | [25] |
pEXP5-CT | General vector containing ampicillin resistance used during sub-cloning to make pACBSR-Hyg | Life Technologies |
pSecTag/FRT/V5-His | Vector containing a promoter-less hygromycin resistance gene | Life Technologies |
pEXP5-CT-hyg | Same as pEXP5-CT, but with the ampicillin resistance marker replaced with a hygromycin resistance marker | This study |
pCP20 | Plasmid bearing a heat-shock inducible FLP recombinase | [26] |
pIJ773 | Template for amplification of the apramycin resistance cassette | [27] |
pACBSR-Hyg | Same as pACBSR, but with the chloramphenicol resistance marker replaced with a hygromycin resistance marker | This study |
pFLP-Hyg | Same as pCP20, but with the pSC101 replicon and chloramphenicol resistance marker of pCP20 replaced with the p15A replicon and hygromycin resistance marker from pACBSR-Hyg | This study |