Natural and synthetic tetracycline-inducible promoters for use in the antibiotic-producing bacteria Streptomyces.
Bacteria in the genus Streptomyces are major producers of antibiotics and other pharmacologically active compounds. Genetic and physiological manipulations of these bacteria are important for new drug discovery and production development. An essential part of any 'genetic toolkit' is the availability of regulatable promoters. We have adapted the tetracycline (Tc) repressor/operator (TetR/tetO) regulatable system from transposon Tn10 for use in Streptomyces. The synthetic Tc controllable promoter (tcp), tcp830, was active in a wide range of Streptomyces species, and varying levels of induction were observed after the addition of 1-100 ng/ml of anhydrotetracycline (aTc). Streptomyces coelicolor contained an innate Tc-controllable promoter regulated by a TetR homologue (SCO0253). Both natural and synthetic promoters were active and inducible throughout growth. Using the luxAB genes expressing luciferase as a reporter system, we showed that induction factors of up to 270 could be obtained for tcp830. The effect of inducers on the growth of S.coelicolor was determined; addition of aTc at concentrations where induction is optimal, i.e. 0.1-1 microg/ml, ranged from no effect on growth rate to a small increase in the lag period compared with cultures with no inducer.
Rodríguez-García
Antonio
A
Institute of Genetics, University of Nottingham, Queens Medical Centre Nottingham NG7 2UH, UK.
Combes
Patricia
P
Pérez-Redondo
Rosario
R
Smith
Matthew C A
MC
Smith
Margaret C M
MC
D000900:Anti-Bacterial Agents / Q000096:biosynthesis; D001483:Base Sequence; D004274:DNA, Recombinant / Q000737:chemistry; D017930:Genes, Reporter; D008969:Molecular Sequence Data; D009875:Operator Regions, Genetic; D011401:Promoter Regions, Genetic; D012097:Repressor Proteins / Q000378:metabolism; D013302:Streptomyces / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D048372:Streptomyces coelicolor / Q000235:genetics / Q000254:growth & development; D013754:Tetracyclines / Q000494:pharmacology*; D015533:Transcriptional Activation
D000900:Anti-Bacterial Agents; D004274:DNA, Recombinant; D012097:Repressor Proteins; D013754:Tetracyclines; C034811:tetracycline resistance-encoding transposon repressor protein; C016229:4-epianhydrotetracycline