Table 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Relevant characteristics | Reference or source |
|---|---|---|
| Strains | ||
| E. coli DH5α | supE44 lacU169 (ϕ80dlacZΔM15) hsdR17 recA1 endA1 gyrA96 Δthi relA1 | 38 |
| E. coli BL21(DE3) | F− ompT hsdSB(rB− mB−) gal dcm lacY1 (DE3) | Novagen |
| E. coli K-12 W3110 | 3HPP utilizer, Kms Tcs | 26 |
| E. coli K-12 W3110ΔmhpT | E. coli K-12 W3110 with the DNA fragment encoding MhpT deleted | This study |
| Plasmids | ||
| pGFPe | Kmr, ColE1 replicon, derivative of pWaldo digested with XbaI and EcoRI, yielding a 14-amino-acid linker sequence (SVPGSENLYFQGQF) followed by GFP | 23 |
| pGFPe-mhpT | PCR-amplified fragment containing mhpT without the stop codon inserted into pGFPe at the XhoI/BamHI restriction sites | This study |
| pGFPe-mhpTE27A | PCR-amplified fragment containing mhpT (mhpTE27A) without the stop codon inserted into pGFPe at the XhoI/BamHI restriction sites | This study |
| pGFPe-mhpTE27D | PCR-amplified fragment containing mhpT (mhpTE27D) without the stop codon inserted into pGFPe at the XhoI/BamHI restriction sites | This study |
| pGFPe-mhpTD75A | PCR-amplified fragment containing mhpT (mhpTD75A) without the stop codon inserted into pGFPe at the XhoI/BamHI restriction sites | This study |
| pGFPe-mhpTD75E | PCR-amplified fragment containing mhpT (mhpTD75E) without the stop codon inserted into pGFPe at the XhoI/BamHI restriction sites | This study |
| pGFPe-mhpTA272H | PCR-amplified fragment containing mhpT (mhpTA272H) without the stop codon inserted into pGFPe at the XhoI/BamHI restriction sites | This study |
| pGFPe-mhpTK276D | PCR-amplified fragment containing mhpT (mhpTK276D) without the stop codon inserted into pGFPe at the XhoI/BamHI restriction sites | This study |
| pVLT31 | Broad-host-range expression vector (IncQ, RSF1010 replicon), Tcr Ptac lacIq | 39 |
| pVLT31-mhpT | PCR fragment containing mhpT inserted into pVLT31 at the EcoRI/HindIII restriction sites | This study |
| pVLT31-mhpTE27A | PCR fragment containing mutant mhpT (mhpTE27A) inserted into pVLT31 at the EcoRI/HindIII restriction sites | This study |
| pVLT31-mhpTE27D | PCR fragment containing mutant mhpT (mhpTE27D) inserted into pVLT31 at the EcoRI/HindIII restriction sites | This study |
| pVLT31-mhpTD75A | PCR fragment containing mutant mhpT (mhpTD75A) inserted into pVLT31 at the EcoRI/HindIII restriction sites | This study |
| pVLT31-mhpTD75E | PCR fragment containing mutant mhpT (mhpTD75E) inserted into pVLT31 at the EcoRI/HindIII restriction sites | This study |
| pVLT31-mhpTA272H | PCR fragment containing mutant mhpT (mhpTA272H) inserted into pVLT31 at the EcoRI/HindIII restriction sites | This study |
| pVLT31-mhpTK276D | PCR fragment containing mutant mhpT (mhpTK276D) inserted into pVLT31 at the EcoRI/HindIII restriction sites | This study |