Structure and Expression of a Chloramphenicol Resistance Gene, cat-86, Cloned From Bacillus Pumilus

The cat-86 gene of Bacillus pumilus, specifying a chloramphenicol-inducible chloramphenicol acetyltransferase was previously cloned and expressed in Bacillus subtilis on plasmid pUB110. The insert DNA has been sequenced and the cat-86 gene identified. The gene encodes a protein 220 amino acids long which has TTG as the translation initiation codon. Putative RNA polymerase and ribosomal binding sites have been identified in sequences at the 5′ end of the gene. Together with a promoter from bacteriophage SP02, the cat-86 gene has been transferred to strains of Bacillus subtilis with ribosomal protein mutations. The influence of these mutations on its expression is described, together with its expression on chimeric plasmids in Escherichia coli.