Identification of Genes Required to Synthesize an Antibiotic-like Compound from the Soil Bacterium Rhodococcus sp. MTM3W5.2

• Main Author: Ward, Amber L
• Format: Others
• Language: English
• Published: Digital Commons @ East Tennessee State University 2015
• Subjects: Rhodococcus; polyketide synthase; antibiotic; natural product; Bacteriology; Microbiology; Other Microbiology
• Online Access: https://dc.etsu.edu/etd/2558; https://dc.etsu.edu/cgi/viewcontent.cgi?article=3935&context=etd

Rhodococcus is a soil bacterium, member of the Actinobacteria, and a close relative of the prolific small molecule producer Streptomyces. Recent interest in Rhodococcus as an under investigated source of possible bioactive secondary metabolites is sparked by the discovery of many polyketide synthase and non-ribosomal peptide synthetase genes of unknown function from sequenced Rhodococcus genomes. Rhodococcus species strain MTM3W5.2 was recently shown to produce a strong inhibitory compound with activity against most strains of Rhodococcus and closely related genera. A goal of this investigation is to discover the gene(s) required to synthesize this inhibitory molecule. The engineered Rhodococcus transposon, pTNR, was used to generate random insertional mutations in the genome of MTM3W5.2. The transposon insertion sites for 8 non-producing mutants were cloned and sequenced. Genes that encode polyketide synthases usually form parts of large biosynthetic gene clusters responsible for the production of small polyketide molecules.