TABLE 1.
Strains of Escherichia coli and plasmids used in this study
| E. coli strain or plasmid | Genotype | Source or reference |
|---|---|---|
| Strains | ||
| MC4100 | F−araD139 Δ(argF-lac)U169 rpsL150 relA1 flB530 deoC1 ptsF25 rbsR | 27 |
| SC1088citA | lacZ4075 mini Tet del citA::Kanr | 13 |
| IMW237 | MC4100, λ[Φ(dcuB′-′lacZ)Hyb Ampr] | 30 |
| IMW260 | MC4100 but λ[Φ(dcuB′-′lacZ)Hyb Ampr] dcuS::Camr | 30 |
| IMW238 | MC4100 but λ[Φ(dcuB′-′lacZ)Hyb Ampr] dcuR::Kanr | 30 |
| IMW280 | MC4100 but λ[Φ(dcuB′-′lacZ)Hyb Ampr] citA::Kanr | IMW237 × P1 (Sc1088citA) |
| IMW239 | MC4100 but λ[Φ(dcuB′-′lacZ)Hyb Ampr] citB::Specr | 30 |
| Plasmids | ||
| pET28a | Novagen | |
| pMW145 | pET28a, comprising the periplasmic domain of DcuS from amino acid Ser45 to Arg180 | 23 |
| pMW423 | pMW145 including periplasmic domain DcuS-PD(T101G F120M I125S)45-180 | This study |
| pMW181 | pET28a with dcuS | 19 |
| pMW345 | pMW181 with dcuS mutant encoding DcuS(F149K) | This study |
| pMW352 | pMW181 with dcuS mutant encoding DcuS(F149K A164S) | This study |
| pMW344 | pMW181 with dcuS mutant encoding DcuS(T101G) | This study |
| pMW368 | pMW181 with dcuS mutant encoding DcuS(F120M) | This study |
| pMW373 | pMW181 with dcuS mutant encoding DcuS(I125S) | This study |
| pMW390 | pMW181 with dcuS mutant encoding DcuS(F120M I125S) | This study |
| pMW351 | pMW181 with dcuS mutant encoding DcuS(T101G F120M) | This study |
| pMW369 | pMW181 with dcuS mutant encoding DcuS(T101G I125S) | This study |
| pMW353 | pMW181 with dcuS mutant encoding DcuS(T101G F120M I125S) | This study |
| pMW354 | pMW181 with dcuS mutant encoding DcuS(F141S) | This study |
| pMW355 | pMW181 with dcuS mutant encoding DcuS(Q159V) | This study |
| pMW356 | pMW181 with dcuS mutant encoding DcuS(Q159V M103A) | This study |
| pMW370 | pMW181 with dcuS mutant encoding DcuS(T101G F120M I125S F149K) | This study |
| pMW374 | pMW181 with dcuS mutant encoding DcuS(T101G F120M I125S A164S) | This study |
| pMW371 | pMW181 with dcuS mutant encoding DcuS(T101G F120M I125S Q159V) | This study |
| pMW372 | pMW181 with dcuS mutant encoding DcuS(T101G F120M I125S F149K A164S) | This study |
| pMW398 | pMW181 with dcuS mutant encoding DcuS(R107A) | This study |
| pMW409 | pMW181 with dcuS mutant encoding DcuS(F149K A164S F141S) | This study |
| pMW410 | pMW181 with dcuS mutant encoding DcuS(T101G F120M I125S F141S) | This study |
| pMW236 | pMW181 with dcuS mutant encoding DcuS(H110A) | This study |
| pMW237 | pMW181 with dcuS mutant encoding DcuS(R147A) | This study |
| pMW277 | pMW181 with dcuS mutant encoding DcuS(F149A) | This study |
| pMW298 | pMW181 with dcuS mutant encoding DcuS(A164S) | |
| pMW296 | pMW181 with dcuS mutant encoding DcuS(M103A) |