Table 1 Bacterial strains and plasmids used in this study
From: Targeted elimination of Vancomycin resistance gene vanA by CRISPR-Cas9 system
Bacterial strains and plasmids | Â | Reference or source |
|---|---|---|
Bacterial strains E. coli DH5α | F-,φ80dlacZΔM15, Δ(lacZYA-argF)U169, deoR, recA1, endA1, hsdR17(rk-, mk+), phoA, supE44, λ-, thi-1, gyrA96, relA1 | This study |
E.coli BL21 | F-, ompT, hsdSB (rB-mB-) gal,dcm araB::T7RNAP-tetA | This study |
VR-E. faecium24 | Enterococcus clinical strains containing vanA | This study |
Plasmids pCas9 (Addgene,42,876) | Cmr, tracRNA, gRNA, and cas9 expression plasmid; | Laboratory stock [15] |
pCas9-sgRNA | Cmr, pCas9 cloned with sgRNA targeting vanA; | This study |
pUC19 (Invitrogen, Carlsbad, CA, USA) | AMPr, expression vector | Laboratory stock |
pUC19-vanA | AMPr, recombinant vector derivative with vanA gene | This study |