Application of transposon mutagenesis to identify global regulatory network in actinomycetes

• Applicant: Professor Dr. Andriy Luzhetskyy
• Subject Area: Metabolism, Biochemistry and Genetics of Microorganisms
• Term: from 2010 to 2013
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 161577302

This project aims to identify gene networks, responsible for the regulation of antibiotic biosynthesis in Streptomyces coelicolor and S. sp. Tü6071, using a random transposon mutagenesis system and fluoroscence-activated cell sorting (FACS) high-throughput screening. The objective is to generate a large repository of actinomycetes events with tagged genes and to isolate mutants with changes in the antibiotic (actinorhodin, prodigiosin and phenalinolactone) production. The changes in antibiotic production will be identified through the expression level of the reporter genes GFP and CFP fused to the different promoters from actinorhodin, prodigiosin and phenalinolactone biosynthetic clusters. The mutants will be characterized by cloning and sequencing of the transposon insertion sites from the chromosomes on the rescue plasmids. The study will lead to the better understanding of the antibiotic biosynthesis regulation and to the rational and faster developing of the antibiotic overproducers. In addition, the outcome of this project is a new platform for the identification of the regulation of the metabolic processes in actinomycetes through transposon mutagenesis, reporter gene expression and FACS high-throughput screening.

• DFG Programme: Research Grants