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AMB Express
Fig. 1 | AMB Express

Fig. 1

From: Construction and application of a CRISPR/Cas9-assisted genomic editing system for Corynebacterium glutamicum

Fig. 1

Construction of CRISPR/Cas9-assisted system. a The overview diagram of vectors pCCG1, pCCG2 and pBS-sgRNA construction, E. coli—C. glutamicum shuttle expression vector pCCG1 and pCCG2 is kanamycin resistance, Cas9 nuclease gene is under the control of the inducible promoter Ptrc, pBL1TS is temperature-sensitive replicon in C. glutamicum, constitutive promoter PtacM and multiple cloning site (MCS) were artificial synthesis, sgRNA were expressed by promoter PtacM. Small sgRNA sequence was ligated with plasmid pBluescript II SK. sgRNA fragment were amplified from the plasmid pBS-sgRNA when needed. b The sequence of multiple cloning sites, including BamHI, NheI, Asc I, AflII and SmaI (XmaI) restriction enzyme sites. c The full nucleotide sequence of sgRNA fragment (171 bp) in plasmid pBS-sgRNA, the red part is Cas9 nuclease recognition site (82 bp)

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