Application of redD, the transcriptional activator gene of the undecylprodigiosin biosynthetic pathway, as a reporter for transcriptional activity in Streptomyces coelicolor A3(2) and Streptomyces lividans.
redD encodes the transcriptional activator of the biosynthetic pathway for undecylprodigiosin, a red-pigmented, mycelium-bound antibiotic made by Streptomyces coelicolor A3(2) and Streptomyces lividans. A promoterless version of redD preceded by the efficiently used tuf1 ribosome binding site was inserted into two different plasmid vectors, providing a convenient reporter of transcriptional activity in both species. One plasmid, plJ2587, replicates autonomously in both Escherichia coli and streptomycetes, while the other, plJ2585, replicates in E. coli and can be transferred to streptomycetes by conjugation or transformation, whereupon it integrates stably at the chromosomal attachment site for the temperate phage phiC31. The utility of the plasmids in detecting not only transcriptional activity, but also its regulation, was confirmed using the rrnAp, ermEp*, and glnRp promoters. The ability to screen visually and spectrophotometrically for red pigmentation should make the vectors particularly attractive for analysing the regulation of gene expression, and for the isolation of mutants, in both S. coelicolor and S. lividans.
van Wezel
G P
GP
Department of Biochemistry, Leiden University, The Netherlands.
White
J
J
Hoogvliet
G
G
Bibb
M J
MJ
D000595:Amino Acid Sequence; D000900:Anti-Bacterial Agents / Q000096:biosynthesis*; D001426:Bacterial Proteins / Q000235:genetics; D001483:Base Sequence; D003001:Cloning, Molecular; D004268:DNA-Binding Proteins; D004926:Escherichia coli; D017930:Genes, Reporter; D008969:Molecular Sequence Data; D010957:Plasmids; D011353:Prodigiosin / Q000031:analogs & derivatives / Q000096:biosynthesis*; D011401:Promoter Regions, Genetic; D015183:Restriction Mapping; D013302:Streptomyces / Q000235:genetics* / Q000378:metabolism*; D015534:Trans-Activators / Q000737:chemistry / Q000235:genetics*; D014158:Transcription, Genetic
D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; C082451:GlnR protein, Streptomyces coelicolor; D015534:Trans-Activators; C077706:redD protein, Streptomyces coelicolor; C047347:undecylprodigiosin; D011353:Prodigiosin