Table 1 Strains and plasmids used in this work
Strains and plasmids | Relative characteristics | Resources |
|---|---|---|
Strains | ||
ATCC8739 | Wild type | Lab collection |
CAR005 | ATCC 8739, M1-37::dxs, M1-46::idi M1-93::Crt, M1-46::SucAB, M1-46::sdh, M1-46::talB | Zhao et al. [11] |
CAR015 | CAR005, ispG-mRSL-4, ispH-mRSL-14 | Lab collection |
CAR025 | CAR015, replacing the promoter of crtEYIB with Ptrc promoter | Lab collection |
CAR025-CrtW | CAR025 ( pSC101-CrtW,pGlpF-dCas9, pgRNA-pSC101bb) | This work |
CAR025-CrtW-CK | CAR025 (pSC101-CrtW,pGlpF-dCas9, pgRNA-N20) | This work |
CAR025-CrtZ | CAR025 (pSC101-CrtZ,pGlpF-dCas9, pgRNA-pSC101bb) | This work |
CAR025-CrtZ-CK | CAR025 (pSC101-CrtZ,pGlpF-dCas9, pgRNA-N20) | This work |
CAR025-CrtZW | CAR025 (pSC101-CrtZW,pGlpF-dCas9, pgRNA-pSC101bb) | This work |
CAR025-CrtZW-CK | CAR025 (pSC101-CrtZW,pGlpF-dCas9, pgRNA-N20) | This work |
8739-RFP | ATCC 8739 (pBAD-RFP, pGlpF-dCas9, pgRNA-RFP) | This work |
8739-RFP-CK1 | ATCC 8739 (pBAD-RFP, pGlpF-dCas9, pgRNA-N20) | This work |
8739-RFP-CK2 | ATCC 8739 (pBAD-RFP, pTrc99A-M, pgRNA-N20) | This work |
Plasmids | ||
pACYC184-M | cat; replace tet with lacI and Ptrc of pTrc99A-M | Zhao et al. [11] |
pCas9 | Cas9 expression plasmid | Zhao et al. (2016) |
pSC101 | low copy plasmid, ori and repA, M1-46 promoter, cat from pACYC184-M2-Pm46 | Lab collection |
pYL501 | crtW and crtZ in pSC101 | Lab collection |
pCrtW | crtW in pSC101 | This work |
pCrtZ | crtZ in pSC101 | This work |
pCrtZW | crtW and crtZ in pSC101 | This work |
pGlpF-dCas9 | GlpF fused with dCas9 in pTrc99A, pMB1 | This work |
pgRNA-RFP | Apr, p15A, gRNA-rfp 20 bp guide sequence(catgcgtttcaaagttcgta) | This work |
pgRNA-pSC101bb | Apr, p15A, gRNA-pYL501bb 20 bp guide sequence(catgtaacacctaatagaac) | This work |
pgRNA-IdhAL-IdhAR | Apr, p15A, gRNA-IdhAL-gRNA-IdhAR 20 bp guide sequence L(cagtaataacagcgcgagaa) 20 bp guide sequence R(tgttgcgctaagcctgctga) | This work |
pBAD-RFP | Kna, pBAD-rfp, repA101 | Lab collection |