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Fig. 1 | Microbial Cell Factories

Fig. 1

From: Combining transposon mutagenesis and reporter genes to identify novel regulators of the topA promoter in Streptomyces

Fig. 1

Scheme of random Tn5 transposon mutagenesis in the S. coelicolor WT-lux reporter strain. 1 Random transposon mutagenesis of the WT-lux strain (MG03) with a mini-Tn5 transposon. 2 The WT-lux-tn mutant library consisted of approximately 8300 single colonies obtained on MS agar plates. 3 Measurement of the luminescence of WT-lux-tn library single colonies. 4 Selection of colonies with altered light emission compared to the WT-lux paternal strain and TopA-depleted lux strain (MG04, high activity of topA promoter). 5 The luminescence of selected colonies from the WT-lux-tn library measured in liquid culture compared with the WT-lux strain and TopA-depleted lux strain (high activity of the topA promoter). RU—relative luminescence units. 6 Western blot analysis of TopA protein level in cell lysates of selected colonies from the WT-lux-tn library with anti-TopA polyclonal antibodies. M—molecular mass marker

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Microbial Cell Factories

ISSN: 1475-2859

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