pKD46

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	The JM109(DE3) cells containing ChroV-GFPuv were grown on LB agar plate containing 25 μg/mL of chloramphenicol and 0.5 mM IPTG at 37°C. The colonies images were detected by colony fluorescent image analysis.

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The JM109(DE3) cells containing ChroV-GFPuv were grown on LB agar plate containing 25 μg/mL of chloramphenicol and 0.5 mM IPTG at 37°C. The colonies images were detected by colony fluorescent image analysis.

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The JM109(DE3) cells containing ChroV-GFPuv were grown on LB agar plate containing 25 μg/mL of chloramphenicol and 0.5 mM IPTG at 37°C. The colonies images were detected by colony fluorescent image analysis. The colonies were transferred into 50 mL of Luria–Bertani (LB) medium in a 250 mL flask containing 25 μg/mL of chloramphenicol at 37°C, induced with 0.5 mM IPTG at three different optical densities at 600 nm, and cultured 37°C for 12 h. Cultured cells were harvested, washed in PBS (pH 7.4), lysed using cell lytic B buffer (Sigma), and analyzed by SDS-PAGE. The thickness and intensities of GFPuv band were analyzed using an open source image processing program, ImageJ [20]. ChroV-JM109(DE3) cells harboring various gene clusters such as GFPuv (0.7 kb), EGFP fused with a maltose binding protein (2 kb), and carotenoid-biosynthetic pathway gene clusters (4 kb, 6 kb, and 10 kb) as described previously [19], were grown at the same conditions containing 0.5 mM IPTG. These large genes and clusters were selected to facilitate the detection of protein expression based on fluorescence- or color-observation of the resulting colonies. Colonies expressing β-carotene and astaxanthin were yellow and red in color, respectively.

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