Relatively limited information about promoter structures in has been available until now. With the aim of isolating and characterizing such transcription initiation signals, random 3A fragments of chromosomal DNA and of the corynebacterial phage øGA1 were cloned into the promoter probe vector pEKplCm and selected for promoter activity by chloramphenicol resistance of transformed cells. The nucleotide sequence of ten chromosomal and three phage fragments was determined and the transcriptional start (TS) sites were localized by primer extension analyses. Additionally, the promoters of five previously isolated genes were cloned and mapped. All of the isolated promoters were also functional in the heterologous host A comparative analysis of the newly characterized promoter sequences together with published promoters from C. revealed conserved sequences centred about 35 bp (ttGcca) and 10 bp (TA.aaT) upstream of the TS site. The position of these motifs and the motifs themselves are comparable to the −35 and −10 promoter consensus sequences of other Gram-positive and Gram-negative bacteria, indicating that they represent transcription initiation signals in However, the consensus hexamer of the −35 region is much less conserved than in and


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