ISOLATION OF PROMOTERS FROM BREVIBACTERIUM-FLAVUM STRAIN MJ233C AND COMPARISON OF THEIR GENE-EXPRESSION LEVELS IN B-FLAVUM AND ESCHERICHIA-COLI

A promoter probe shuttle vector suitable for the isolation of promoter elements from coryneform bacteria was constructed. This vector carrie d the neomycin phosphotransferase (NPTII) gene from transposon Tn5 as a reporter gene, and was capable of replication in both Escherichia co li and Brevibacterium flavum. The vector was used in the construction of a B. flavum library of 899 independently isolated promoter clones. Promoters with a wide range of activities in B. flavum, including some very strong promoter elements, were isolated. Comparative analysis su ggests that significant differences between B. flavum and E. coli may exist in the determinants of promoter strength.