TABLE 1.
E. coli K12 strains and plasmids
| Strain or plasmid | Genotype and/or properties (reference); source |
|---|---|
| Strains | |
| PC2 | leuB6 thyA47 dnaC2 dnaT12 deoC3 rpsL153 λ− (5); laboratory stock |
| SS1020 | λ−rph-1 zjj202::Tn10 dnaC2; Steven Sandler |
| HME5 | F− λ− IN (rrnD-rrnE) ΔlacU169 λcI857 Δ(cro-bioA) (32); Donald Court |
| MC1061 | araD139 Δ(ara, leu)7697 ΔlacX74 galU galK rpsL hsdR2 (rK− mK+) mcrB1; laboratory stock |
| KH1061 | araD139 Δ(ara, leu)7697 ΔlacX74 galU galK rpsL hsdR2 (rK− mK+) mcrB1 ΔdnaC::cat; this work |
| MF1061 | araD139 Δ(ara, leu)7697 ΔlacX74 galU galK rpsL hsdR2 (rK− mK+) mcrB1 ΔdnaC::cat recA::Kanr; Magdalena M. Felczak of this laboratory by P1 transduction from BW26355 (relevant genotype, ΔrecA635::Kanr) (8) into MC1061, followed by selection for kanamycin resistance; the transductant and BW26355 were confirmed to be UV sensitive |
| Plasmids | |
| pACYC184 | cat, Tetr (6) |
| pACYC184dnaC | Tetr, dnaC (this work) |
| pAM34 | Ampr, IPTG-dependent maintenance (32); Kaymeuang Cam |
| pAM34dnaCL1-2 | Ampr, IPTG-dependent maintenance, dnaC (this work) |
| pAM34dnaCL1-4 | Ampr, IPTG-dependent maintenance, dnaC (this work) |
| pINCSSD | Kanr, pBR322 derivative, dnaC under araBAD promoter control and downstream from the ribosome binding site for gene 10 of T7 phage (1) |
| pdnaC113 | Ampr, pBR322 derivative, dnaC under araBAD promoter control (20) |
| pdnaC97 (Pro108Ser) | Kanr, pINCSSD derivative, missense mutation encoding Pro108Ser substitution (20) |
| pdnaC116 (Leu44Pro) | Kanr, pINCSSD derivative, missense mutation encoding Leu44Pro substitution (20) |
| pdnaC250 (Leu11Gln) | Kanr, pINCSSD derivative, missense mutation encoding Leu11Gln substitution (20) |
| pHK-dnaC | Ampr, pET11a derivative (Novagen), dnaC joined at the 5′ end to sequence encoding protein kinase A phosphorylation site and six histidine residues (9); Mike O'Donnell |
| pHK-dnaCK112R | Ampr, derivative of pHK-dnaC encoding a Lys112Arg substitution (9); Mike O'Donnell |