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Fig. 5 | Microbial Cell Factories

Fig. 5

From: Comparative genomics and transcriptomics analyses provide insights into the high yield and regulatory mechanism of Norvancomycin biosynthesis in Amycolatopsis orientalis NCPC 2-48

Fig. 5

AoStrR1 binds to the promoter regions of the nvcm cluster. a Overexpression and purification of His10-tagged AoStrR1 in E. coli. SDS-PAGE showing purification of N-terminal His10-tagged AoStrR1 by nickel-affinity chromatography. lanes 1–5, elution with 500 mM imidazole; lanes 6–7, desalted by the PD10 Desalting Columns. M stands for protein size marker. b AoStrR1 binds to the 8 promoter regions identified by EMSA. Lane −, probe only; Lane +, 1,000 nM His10-AoStrR1 incubated with the probe; Lane C, 1000 nM His10-AoStrR1 incubated with 100-fold excess unlabeled specific competitor DNA fragment. c Sequence alignment of the putative AoStrR1 binding sites. Consensus sequence was generated using MEME. Arrows indicate inverted repeats and the double-headed arrow denotes the spacer. The most highly conserved nucleotides are bold. d The genes present in norvancomycin BGC are indicated by arrows. The short bars are the probes in the nvcm cluster. Red triangles are the binding sites of His10-AoStrR1

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Microbial Cell Factories

ISSN: 1475-2859

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