Table 2 Bacterial strains and plasmids used in this study
From: Role of AcsR in expression of the acetyl-CoA synthetase gene in Vibrio vulnificus
Strain/plasmid | Genotype | Source/Reference |
|---|---|---|
Escherichia coli | ||
DH5α | (Φ80 lacZ ΔM15) recA1 endA1 gyrA96 relA1 thi-1 hsdR17(rK − mK −) supE44 deoR (lacZYA-argF)U169 | Invitrogen |
BL21(DE3) | E. coli strain B F− dcm ompT hsdS(rB − mB −) galλ(DE3)thi-1 thr leu tonA lacY supE recA:: | Invitrogen |
SM10λpir | Rp4-2-Tc::Muλpir, oriT of RP4, Kmr endA1 recA1 gyrA96 thi-1 hsdR17(rK − mK −) relA1supE44 Δ(lac-proAB)[F’traD36proABlacI qZΔM15] | [55] |
Vibrio vulnificus | ||
MO6-24/O | Clinical isolate | [56] |
MJ1 | MO6-24/O, ΔvarA | This study |
MJ2 | MO6-24/O, ΔacsR | This study |
MJ3 | MO6-24/O, ΔacsS | This study |
MJ4 | MO6-24/O, ΔacsA | This study |
KJLΔcya | MO6-24/O, Δcya | This study |
Plasmids | ||
pBluescript(II)SK(+) | Cloning vector; Apr, lac promoter (lacZ), f1, ColE1 | Stratagene |
pSKvarAU | pBluescript(II)SK(+) with 501-bp upstream region of varA | This study |
pSKvarAUD | pSKvarAU with 520-bp downstream region of varA | This study |
pSKacsRU | pBluescript(II)SK(+) with 410-bp upstream region of acsR | This study |
pSKacsRUD | pSKacsRU with 424-bp downstream region of acsR | This study |
pSKacsSU | pBluescript(II)SK(+) with 710-bp upstream region of acsS | This study |
pSKacsSUD | pSKacsSU with 528-bp downstream region of acsS | This study |
pSKacsAU | pBluescript(II)SK(+) with 732-bp upstream region of acsA | This study |
pSKacsAUD | pSKacsAU with 525-bp downstream region of acsA | This study |
pDM4 | Suicide vector; oriR6K, Cmr | [57] |
pDMΔcya | pDM4 containing 1,272-bp DNA of internally deleted cya gene | [41] |
pDMΔvarA | pDM4 containing 1,021-bp SalI/XbaI fragment of pSKvarAUD | This study |
pDMΔacsR | pDM4 containing 834-bp ApaI/sacI fragment of pSKacsRUD | This study |
pDMΔacsS | pDM4 containing 1,238-bp XhoI/XbaI fragment of pSKacsSUD | This study |
pDMΔacsA | pDM4 containing 1,953-bp XhoI/XbaI fragment of pSKacsAUD | This study |
pHK0011 | pRK415, a promoterless luxAB, Tcr | [58] |
pHKacsA::luxAB | pHK0011 vector containing 284-bp acsA promoter | This study |
pRK415 | A broad-host-range vector; oriT of RP4; Tcr | [59] |
pRKacsR | pRK415 containing636-bp V. vulnificus acsR ORF | This study |
pRKacsS | pRK415 containing3,432-bp V. vulnificus acsS ORF | This study |
pRKacsA | pRK415 containing1,953-bp V. vulnificus acsA ORF | This study |
pET28b(+) | Expression vector; T7 lac promoter, oriF1; Kmr | Novagen |
pETacsR | pET28b (+) vector containing 636-bp acsR | This study |