HTS of Halomonas: pMT23opt-MmP1.R1
To characterize the orthogonal transcription mutators, three dual-type mutators (pMT23d-MmP1, pMT32d-MmP1, pMT23opt-MmP1) and control pMT0-MmP1 were introduced into H. bluephagenesis strains harboring the genomic sacB gene. Strains were cultured in 60LB medium with 200 mg/L IPTG at 37C for 20 h for mutagenesis. A 3585-bp sacB fragment (containing 2024-bp target region with flanking sequences) was PCR-amplified and subjected to Illumina PE150 sequencing (GENEWIZ, China).