Table 1 Bacterial strains and plasmids used in this study
From: CRISPR/Cas9-deaminase enables robust base editing in Rhodobacter sphaeroides 2.4.1
Strains/plasmids | Characteristics | Source |
|---|---|---|
Rhodobacter sphaeroides 2.4.1 | Wild type | ATCC 17023 |
Rhodobacter sphaeroides KD131 | Wild type | This lab |
Escherichia coli Trans1-T1 | F−φ80(lacZ)ΔM15ΔlacX74hsdR(r −k , m +k )ΔrecA1398endA1tonA | Transgene CD501-01 |
pACYC-Cas9(dCas9/nCas9D10A) | Derive from pACYCDuet-1, CmR, insert Cas9 or its variant in MCS1 | This lab |
pSCI_dCas9-CDA1-UL | Expression plasmid, CmR, contain dCas9, CDA1, UGI, and LVA tag | Gift from Prof. Satomi Banno et al. |
pgRNA-bacteria | Derive from pUC19, AmpR, contain sgRNA scaffold | Addgene 44251 |
pMV-Tada-TadA*(opt) | Gene synthetic plasmid, AmpR, contain codon-optimized TadA–TadA*(opt) | Wuxi Qinglan Biotechnology Inc. |
pIND4 | Expression plasmid, pMB1 ori, LacI, KanaR | Gift from Prof. Judith P.Artimage |
pK18mobSacB | Suicide plasmid, pMB1 ori, SacB, KanaR | This lab |
pIND4-SacB-MCS | Derive from pIND4, insert multiple cloning site(MCS)and SacB | This study |
pIND4-Cas9-sgRNA | Derive from pIND4-SacB-MCS, insert Cas9 and sgRNA | This study |
pIND4-Cas9-sgRNA-up-dw | Derive from pIND4-SacB-MCS, insert Cas9, sgRNA, and homologous arms | This study |
pIND4-dCas9(nCas9D10A)-CDA1-UL-sgRNA | Derive from pIND4-SacB-MCS, abbreviated as d/nCBE-sgRNA | This study |
pIND4-TadA–TadA*(opt)-dCas9(nCas9D10A)-sgRNA | Derive from pIND4-SacB-MCS, abbreviated as d/nABE-sgRNA | This study |