Workflow of single-cell quantification of filamentous fungal promoters to discover cryptic natural products. (IMAGE)
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(A) Fungal promoter library was generated on the basis of the selected promoter sequences from the Aspergillus Genome Database. (B) Protoplasts were prepared in a 24-well microtiter plate and collected for high-throughput flow cytometry (FC) detection in a 96-well microtiter plate. (C) FC-based single-cell quantitative method was established for the detection of gene expression in A. nidulans. (D) The screened promoters were applied for silent biosynthetic gene activation and novel structure discovery. (E) Relative strengths of 43 fungal promoters with >1.5-fold expression level over the commonly used PgpdA.
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