Fig. 3.

dCas9 mediated deregulation of ilvE gene (A) Schematic diagram of ilvE promoter from EcoCyc E. coli database showing spacer sequences within promoter IlvE (p) and gene region IlvE (g) designed for dCas9 targeting. (B) Colony PCR for screening of transformed colonies for successful integration of spacer regions into dCas9. L, represents 10 kb ladder; expected sizes for correct integration are represented by arrows. PCR products are separated on 1.5% agarose gel. ‘C’ represents control. (C) Effect of dCas9-mediated downregulation at the ORF using dcas9IlvE (g) primer (MG02) and promoter region using dcas9IlvE (p) primer (MG03) on the growth of E. coli having KIV pathway integrated. Data is replicate of three values. Error bars represent standard deviation.