TABLE 1.
Plasmids and strains used in this work
| Plasmid or strain | Relevant genotype and/or characteristic(s) | Source or reference |
|---|---|---|
| Plasmids | ||
| pMD19-T vector | TA cloning | TaKaRa |
| pKD13 | R6KY ori, Kanr, Ampr, Cmr | CGSC |
| pKD46 | Ampr, helper plasmid | CGSC |
| pCP20 | Ampr, Cmr, helper plasmid | CGSC |
| pETDuet | T7 promoters, pBR322 ori, Ampr | Novagen |
| pET28a | T7 promoters, pBR322 ori, Kanr | Novagen |
| pETDuet-gfp | pETDuet derivate with gfp cloned | This work |
| pET28a-amy | pET28a derivate with amy cloned | This work |
| Gene knockout cassettes | ||
| ΔdacA::kan | Kanr, knockout of gene dacA | This work |
| ΔdacA::kan′ | Kanr, knockout of gene dacA of BL21 ΔdacB to obtain double deletion mutant | This work |
| ΔdacB::kan | Kanr, knockout of gene dacB | This work |
| Strains | ||
| E. coli JM109 | Cloning host | Novagen |
| E. coli BL21 | Wild-type E. coli BL21(DE3) | Novagen |
| BL21-pETDuet | E. coli BL21 with plasmid pETDuet | This work |
| BL21-pETDuet-gfp | E. coli BL21 with plasmid pETDuet-gfp | This work |
| BL21-pETDuet-amy | E. coli BL21 with plasmid pETDuet-amy | This work |
| BL21-pKD46 | E. coli BL21(DE3) derivate, including plasmid pKD46, Ampr | This work |
| BL21 ΔdacA::kan pKD46 | BL21-pKD46 derivate, deleting dacA | This work |
| BL21 ΔdacB::kan pKD46 | BL21-pKD46 derivate, deleting dacB | This work |
| BL21 ΔdacA ΔdacB::kan pKD46 | BL21 ΔdacB pKD46 derivate, deleting ΔdacA ΔdacB | This work |
| BL21 ΔdacA::kan | BL21 ΔdacA::kan pKD46 derivate, deleting plasmid pKD46 | This work |
| BL21 ΔdacB::kan | BL21 ΔdacB::kan pKD46 derivate, deleting plasmid pKD46 | This work |
| BL21 ΔdacA ΔdacB::kan | BL21 ΔdacA ΔdacB::kan pKD46 derivate, deleting plasmid pKD46 | This work |
| BL21 ΔdacA | BL21 ΔdacA::kan derivate, deleting plasmid pKD46 and Kanr | This work |
| BL21 ΔdacB | BL21 ΔdacB::kan derivate, deleting plasmid pKD46 and Kanr | This work |
| BL21 ΔdacA ΔdacB | BL21 ΔdacA ΔdacB::kan derivate, deleting plasmid pKD46 and Kanr | This work |
| BL21 ΔdacA pETDuet | BL21 ΔdacA derivate with plasmid pETDuet | This work |
| BL21 ΔdacB pETDuet | BL21 ΔdacB derivate with plasmid pETDuet | This work |
| BL21 ΔdacA ΔdacB pETDuet | BL21 ΔdacA ΔdacB derivate with plasmid pETDuet | This work |
| BL21 ΔdacA pETDuet-gfp | BL21 ΔdacA derivate with plasmid pETDuet-gfp | This work |
| BL21 ΔdacB pETDuet-gfp | BL21 ΔdacB derivate with plasmid pETDuet-gfp | This work |
| BL21 ΔdacA pETDuet-amy | BL21 ΔdacA derivate with plasmid pETDuet-amy | This work |
| BL21 ΔdacB pETDuet-amy | BL21 ΔdacB derivate with plasmid pETDuet-amy | This work |
| BL21 ΔdacA ΔdacB pETDuet-gfp | BL21 ΔdacA ΔdacB derivate with plasmid pETDuet-gfp | This work |
| BL21 ΔdacA ΔdacB pETDuet-amy | BL21 ΔdacA ΔdacB derivate with plasmid pETDuet-amy | This work |