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. Author manuscript; available in PMC: 2020 Oct 1.
Published in final edited form as: J Ind Microbiol Biotechnol. 2019 May 15;46(9-10):1359–1364. doi: 10.1007/s10295-019-02187-1

Fig. 2.

Fig. 2

Cloning and heterologous expression of the daq biosynthetic gene cluster. (a) Schematic representation of the in vitro cloning strategy. The daq BGC was PCR-amplified in two fragments of ~12 kb and assembled in vitro using NEBuilder. (b) Restriction digest and gel electrophoresis of plasmids isolated from five different clones. Clones #1–4 were confirmed as positive, and the plasmid was named pJB038EL. L, DNA molecular ladder. (c) HPLC analysis of culture extracts with detection at 280 nm. The extract from the S. coelicolor / pJB038EL exconjugant is ten-fold more concentrated than the extract from the F001 wild-type strain. DAQ analogs produced are indicated. (d) Overlaid UV- Vis spectra of DAQ peaks shown in panel c and a DAQ A standard