Table 1 Strains and plasmids used in the study
From: Pyruvate kinase is necessary for Brucella abortus full virulence in BALB/c mouse
Strains and plasmids | Characteristics | Source |
|---|---|---|
Brucella abortus | ||
S2308 | Wild type strain; Smooth phenotype | ATCC |
∆pyk | pyk gene deletion mutant strain; Smooth phenotype | This study |
∆pyk(Pyk-3 × Flag) | Cmr; complementation strain; ∆pyk carrying the complementation plasmid pBBR-pyk-3 × Flag; Smooth phenotype | This study |
Escherichia coli | ||
DH5α | F− φ80lacZ∆M15 ∆(lacZYA-argF)U169 recA1 endA1 hsdR17(r −k , m +k ) phoA supE44 thi-1 gyrA96 relA1 λ− | Invitrogen |
Plasmids | ||
pBBR1MCS1 | Cmr; Broad-host-range cloning vector | [17] |
pSC | Ampr; pUC19 plasmid containing SacB gene | [16] |
pSC-∆pyk | Ampr; pSC plasmid containing the ∆pyk fragment; used to construct deletion strain | This study |
p3 × Flag-CMV-14 | Ampr; eukaryotic expression plasmid | Sigma-Aldrich |
pBBR-pyk-3 × Flag | Cmr; pBBR1MCS1 containing the pyk gene flanked by its upstream and downstream regions containing a C-terminal 3 × flag tag. | This study |