Table 3.
Strains and plasmids used in this study
| Name | Relevant genotype | Source |
|---|---|---|
|
Strains |
|
|
|
E. coli JM109 |
recA1, endA1, gyrA96, thi-1, hsdR17, supE44, relA1, Δ(lac-proAB)/F’[traD36, proAB+, lacIq, lacZΔM15] |
TransGen Biotech |
| BSUL03 |
ΔamyE::(P43::kivd-adh2), P43::ilvD-ilvC-alsS; Spcr, Emr |
[6] |
| BSUL04 |
BSUL03 with lactate dehydrogenase inactivation (Δldh); Spcr, Emr, Kmr |
This study |
| BSUL05 |
BSUL04 with pyruvate dehydrogenase complex E2 subunit inactivation (ΔldhΔpdhC); Spcr, Emr, Kmr, Tetr |
This study |
|
Plasmids |
|
|
| pUC18 |
E. coli cloning vector; Ampr |
Laboratory stock |
| pDK |
B. subtilis integration vector with amyE locus; Ampr, Kmr |
BGSC [50] |
| pHY300PLK |
E. coli-B. subtilis cloning vector; Ampr, Tetr |
Takara |
| pUCL |
pUC18 containing ldh fragment of B. subtilis, Ampr |
This study |
| pUCLKm |
pUCL containing kanamycin resistance cassettes from pDK; Ampr, Kmr |
This study |
| pUCP01 |
pUC18 containing homology arm pdhC1 fragment of B. subtilis; Ampr |
This study |
| pUCP02 |
pUCP01 containing homology arm pdhC2 fragment of B. subtilis; Ampr |
This study |
| pUCPTet | pUCP02 containing tetracycline resistance cassettes from pHY300PLK; Ampr, Tetr | This study |