Construction and application of streptomycete promoter probe vectors which employ the Streptomyces glaucescens tyrosinase-encoding gene as reporter

Abstract

We have constructed multicopy and integrative streptomycete promoter probe vectors that employ the promoterless tyrosinase ( melC) operon of Streptomyces glaucescens as the chromogenic transcriptional reporter. Each vector contains the reporter cassette, RC3, which comprises part of the melC operon flanked by transcription terminators; RC3 may be easily inserted into any vector. We demonstrate the use of the pIJ101-based mel vector, pMT3010, for the isolation of mutations which alter expression of the S. coelicolor glycerol operon. The S. glaucescens mel reporter system is well-suited for the visual, non-selective identification of regulatory mutants and can be used efficiently for screening several thousand clones at high colony density.