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Figure 1 | Microbial Cell Factories

Figure 1

From: Dissecting and engineering of the TetR family regulator SACE_7301 for enhanced erythromycin production in Saccharopolyspora erythraea

Figure 1

Inactivation of SACE_7301 in S. erythraea A226. (A) Schematic inactivation of SACE_7301 in A226 by linearized fragment homologous recombination. (B) Confirmation of the ΔSACE_7301 mutant by PCR analysis using the primers 7301-P1 and 7301-P4. Lanes: M, 5000-bp DNA ladder. The size of 3,660 bp for a PCR-amplified band was detected in A226, while the bands of the size 4,360 bp were observed in pUCTSR-Δ7301 and ΔSACE_7301, indicating that SACE_7301 was replaced with tsr. (C) Inhibition tests of A226, ΔSACE_7301 and ΔbldD fermentation broths against B. subtilis PUB110. (D) Time course of Er-A yield in A226 and ΔSACE_7301 by HPLC analysis; Mean values of at least three replicates were shown, with the standard deviation indicated by error bars. (E) Growth curves of A226 and ΔSACE_7301. The two strains were cultured in the R5 liquid fermentation medium, and their dry weights of mycelia (DWM) were measured. (F) Er-A production in A226, ΔSACE_7301, ΔSACE_7301/pZMW7301, A226/pZMW, and A226/pZMW7301 cultured in R5 liquid fermentation medium for 6 days by HPLC analysis. Mean values of at least three replicates were shown, with the standard deviation indicated by error bars.

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