From: Development of a CRISPR/Cas9 genome editing toolbox for Corynebacterium glutamicum
Strain or plasmid | Descriptiona | Reference or source |
---|---|---|
E. coli | ||
 DH5α | General cloning host | TaKaRa |
 MG1655 | Source of recT | Lab collection |
C. glutamicum | ||
 ATCC 13869 | Wild-type strain | ATCC |
 ATCC 13032 | Wild-type strain | ATCC |
 ATCC 13032::rfp | ATCC 13032 derivative with insertion of a rfp cassette | Lab collection |
 SL4 | ATCC 13869 derivative with high electrotransformation efficiency | Lab collection |
 SL4 (pCas9) | SL4 derivative harboring pCas9 | This work |
 SL4ΔldhA | SL4 derivative with ldhA deletion | This work |
 SL4ΔldhA::rfp | SL4 derivative with insertion of a rfp cassette in ldhA | This work |
 SL4ΔldhA::rfp (pgRNA5) | SL4ΔldhA::rfp derivative harboring pgRNA5 | This work |
 SL4ΔldhA::rfp off1 | SL4ΔldhA::rfp derivative with nonsense mutation in rfp, constructed by ssDNA (rfp-off1) recombineering | This work |
 SL4 (pgRNA6) | SL4 derivative harboring pgRNA6 | This work |
 SL4 rpsL K43R | SL4 derivative with K43R mutation of rpsL | This work |
 SL4 (pcas9 + pgRNA1) | SL4 derivative harboring pcas9 and pgRNA1 | This work |
 SL4 (pcas9 + pgRNA2) | SL4 derivative harboring pcas9 and pgRNA2 | This work |
 ATCC 13869ΔldhA | ATCC 13869 derivative with ldhA deletion | This work |
 ATCC 13032ΔldhA | ATCC 13032 derivative with ldhA deletion | This work |
 ATCC 13032Δcgl1776-cgl178 | ATCC 13032 derivative with cgl1776-cgl178 deletion | This work |
 ATCC 13032::lacZ | ATCC 13032 derivative with lacZ insertion | This work |
 ATCC 13032::rfp (pgRNA5) | ATCC 13032::rfp derivative harboring pgRNA5 | This work |
 ATCC 13032::rfp off1 | ATCC 13032::rfp derivative with nonsense mutation in rfp, constructed by ssDNA (rfp-off1) recombineering | This work |
 ATCC 13032::rfp off2 | ATCC 13032::rfp derivative with nonsense mutation in rfp, constructed by ssDNA (rfp-off2) recombineering | This work |
Plasmid | ||
 pEC-XK99E | Expression vector of C. glutamicum, IPTG-inducible promoter P trc , KmR | [55] |
 pXMJ19 | Expression vector of C. glutamicum, IPTG-inducible promoter P tac , CmR | [56] |
 pRed_Cas9_recA_Δpoxb300 | Source of cas9 gene and gRNA fragment | [54] |
 pEC-XK99E-rfp | Source of rfp gene | Lab collection |
 pCas9 | pXMJ19 derivative carrying cas9 gene, driven by IPTG-inducible promoter P tac | This work |
 pRfp1 | pXMJ19 derivative carrying cas9 180bp-rfp artificial gene, driven by IPTG-inducible promoter P tac | This work |
 pRfp2 | pXMJ19 derivative carrying cas9 180bp-rfp artificial gene, driven by propionate-inducible promoter P prpD2 | This work |
 pRfp3 | pEC-XK99E derivative carrying gRNA-rfp cassette, driven by constitutive promoter P 11F | This work |
 pRfp4 | pEC-XK99E derivative carrying gRNA (with T sp deleted)-rfp cassette, driven by constitutive promoter P 11F | This work |
 pgRNA1 | pEC-XK99E derivative carrying gRNA targeting ldhA, driven by IPTG-inducible promoter P tac | This work |
 pgRNA2 | pEC-XK99E derivative carrying gRNA targeting ldhA, driven by constitutive promoter P 11F , followed by T rrnB without the 185 bp redundant segment | This work |
 pgRNA3 | pgRNA2 derivative carrying homologous arms (~ 1 kb) for ldhA deletion and corresponding gRNA cassette | This work |
 pgRNA4 | pgRNA3 derivative with a rfp cassette inserted between the two homologous (~ 1 kb) arms for replacing ldhA with the rfp cassette | This work |
 pgRNA5 | pgRNA2 derivative carrying gRNA targeting rfp and a P prpD2 -recT cassette for ssDNA-directed rfp editing | This work |
 pgRNA6 | pgRNA5 derivative carrying gRNA targeting rpsL and a P prpD2 -recT cassette for ssDNA-directed rpsL editing | This work |
 pgRNA7 | pgRNA5 derivative carrying two gRNAs targeting rfp and rpsL and a P prpD2 -recT cassette for double-locus editing | This work |
 pgRNA8 | pgRNA3 derivative carrying homologous arms (~ 0.5 kb) for ldhA deletion and corresponding gRNA cassette | This work |
 pgRNA9 | pgRNA3 derivative carrying homologous arms (~ 1 kb) for cgl1776-cgl1781 deletion (8083 bp) and corresponding gRNA cassette | This work |
 pgRNA10 | pgRNA3 derivative carrying homologous arms (~ 1 kb), lacZ cassette (3626 bp) and corresponding gRNA cassette for lacZ cassette insertion | This work |