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Bioresources and Bioprocessing
Fig. 1 | Bioresources and Bioprocessing

Fig. 1

From: Development of chromosome-based T7 RNA polymerase and orthogonal T7 promoter circuit in Escherichia coli W3110 as a cell factory

Fig. 1

Schematic diagram of the engineered W3110 strain with chromosome-equipped T7RNAP a genetic design of engineered T7RNAP in E. coli W3110. Integration was achieved by site-specific recombination at the HK022 attB site. Three engineered T7RNAP expression strains were created. The strain W3110::IL5 has a lacI-PLacUV5-lacZ’-T7RNAP copy from BL21(DE3), and W3110::L5 lacks an additional integrated lacI. W3110::pI, in which T7RNAP is driven by the PLacI without additional repressor and lacZ’ sequence near the promoter. The characteristic fragments of each strain were labeled by primers and result of confirmation was showed in b: lane 1: PCR with primer lacI-F and T7-sR of W3110::IL5, lane 2: PCR with primer lacZ’-F and T7-sR of W3110::L5, lane 3: PCR with primer PlacI-F and T7-sR of W3110::pI. The desired fragment of lacI-F and T7-sR is 1068 bp, lacZ’-F and T7-sR is 637 bp and 522 bp for PlacI-F and T7-sR. M: 100 bp marker

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