Table 1 E. coli strains and plasmids used in this study
Name | Description or relevant genotype | Source |
|---|---|---|
E. coli host strains | ||
DH5α | F − , endA1, glnV44, thi-1, recA1, relA1, gyrA96, deoR, nupG φ80d lacZ∆acZd ladlacZYA – argF) U169, hsdR17(rK-mK +), λ- | Lab stock |
BW25113 | F-, ∆(araD-araB)567, ∆lacZ4787(::rrnB-3), λ-, rph-1, ∆(rhaD-rhaB)568, hsdR514 | (Datsenko and Wanner 2000) |
BW∆ldhA | BW25113 ldhA null mutant | (Srirangan et al. 2014) |
DMH | BW∆ldhA/pK-hemA | (Miscevic et al. 2021) |
DMH∆sdhA | sdhA null mutant of DMH | (Miscevic et al. 2021) |
DMH∆iclR | iclR null mutant of DMH | This study |
DMH∆iclR∆sdhA | iclR and sdhA mutants of DMH | (Miscevic et al. 2021) |
DMH-D9∆sdhA | DMH∆sdhA/pK-hemA/pgRNA-D9/pdcas9-bacteria | This study |
DMH-D9∆iclR∆sdhA | DMH∆iclR∆sdhA /pK-hemA/pgRNA-D9/pdcas9-bacteria | This study |
DSL | BW∆ldhA/pK-hemA-hemB | This study |
DSL∆sdhA | sdhA null mutant of DSL | This study |
DSL∆iclR | iclR null mutant of DSL | This study |
DSL∆iclR∆sdhA | iclR and sdhA mutants of DSL | This study |
DSL-D9∆sdhA | DSL∆sdhA/pK-hemA-hemB/pgRNA-D9/pdcas9-bacteria | This study |
DSL-D1∆iclR∆sdhA DSL-D2∆iclR∆sdhA DSL-D3∆iclR∆sdhA DSL-D4∆iclR∆sdhA DSL-D5∆iclR∆sdhA DSL-D6∆iclR∆sdhA DSL-D7∆iclR∆sdhA DSL-D8∆iclR∆sdhA DSL-D9∆iclR∆sdhA | DSL∆iclR∆sdhA /pK-hemA-hemB/pgRNA-D1/pdcas9-bacteria DSL∆iclR∆sdhA /pK-hemA-hemB/pgRNA-D2/pdcas9-bacteria DSL∆iclR∆sdhA /pK-hemA-hemB/pgRNA-D3/pdcas9-bacteria DSL∆iclR∆sdhA /pK-hemA-hemB/pgRNA-D4/pdcas9-bacteria DSL∆iclR∆sdhA /pK-hemA-hemB/pgRNA-D5/pdcas9-bacteria DSL∆iclR∆sdhA /pK-hemA-hemB/pgRNA-D6/pdcas9-bacteria DSL∆iclR∆sdhA /pK-hemA-hemB/pgRNA-D7/pdcas9-bacteria DSL∆iclR∆sdhA /pK-hemA-hemB/pgRNA-D8/pdcas9-bacteria DSL∆iclR∆sdhA /pK-hemA-hemB/pgRNA-D9/pdcas9-bacteria | This study This study This study This study This study This study This study This study This study |
Plasmids | ||
pCP20 pK184 | Flp + , λ cI857 + , λ pR Rep(pSC101 ori)ts, ApR, CmR p15A ori, KmR, Plac::lacZ’ | (Cherepanov and Wackernagel 1995) (Jobling and Holmes 1990) |
pdcas9-bacteria | p15A ori, PTet-dCas9 | (Qi et al. 2013) |
pgRNA-bacteria | ColE1 origin, PJ23119-gRNA | (Qi et al. 2013) |
pgRNA-D1 pgRNA-D2 pgRNA-D3 pgRNA-D4 pgRNA-D5 pgRNA-D6 pgRNA-D7 pgRNA-D8 pgRNA-D9 | Derived from pgRNA-bacteria, PspeI::hemC-gRNA-D1 Derived from pgRNA-bacteria, PspeI::hemC-gRNA-D2 Derived from pgRNA-bacteria, PspeI::hemC-gRNA-D3 Derived from pgRNA-bacteria, PspeI::hemC-gRNA-D4 Derived from pgRNA-bacteria, PspeI::hemC-gRNA-D5 Derived from pgRNA-bacteria, PspeI::hemC-gRNA-D6 Derived from pgRNA-bacteria, PspeI::hemC-gRNA-D7 Derived from pgRNA-bacteria, PspeI::hemC-gRNA-D8 Derived from pgRNA-bacteria, PspeI::hemC-gRNA-D9 | This study This study This study This study This study This study This study This study This study |
pK-hemA | Derived from pK184, Plac::hemA | (Miscevic et al. 2021) |
pK-hemA-hemB | Derived from pK184, Plac::hemA-hemB | This study |