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Bioresources and Bioprocessing
Fig. 5 | Bioresources and Bioprocessing

Fig. 5

From: Protein expression analysis of a high-demeclocycline producing strain of Streptomyces aureofaciens and the roles of CtcH and CtcJ in demeclocycline biosynthesis

Fig. 5

Overexpression of the individual ctcH and ctcJ genes and the simultaneous overexpression of ctcH and ctcJ. a Genotypes of WT and overexpressing mutants. b Confirmation of the ctcH-overexpressing, ctcJ-overexpressing, and ctcH–ctcJ-overexpressing mutants with PCR. The PCR products were 900 bp for pIB139 (empty vector), 1663 bp for ctcJ+, 1619 bp for ctcH+, and 2382 bp for ctcH–ctcJ+. All the PCR products were sequenced to confirm each mutation. c 6-DCT production during the fermentation of the parental strain DT1, and the ctcH-overexpressing, ctcJ-overexpressing, and ctcH–ctcJ-overexpressing mutants. d RT-qPCR analysis of ctcH and ctcJ expression in the three overexpressing mutants and WT. Relative levels of ctcH and ctcJ transcripts were determined after normalization to the internal reference, 16S rRNA. RT-qPCR was performed in three independent experiments, each in triplicate

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