Construction of a promoter-probe shuttle vector for Escherichia coli and brevibacteria.

Abstract We constructed a promoter-probe vector, pJUP05, for brevibacteria and Escherichia coli based on the promoterless neomycin-resistance ( neo R ) gene from Tn5. This gene confers resistance to the aminoglycosides, kanamycin and neomycin. The promoter of the neo R gene was deleted and replaced by a suitable multiple cloning site. There are translation stop codons in all three reading frames upstream from the neo R gene. The plasmid contains functional origins of DNA replication for both brevibacteria and E. coli , and permits selection for chloramphenicol- and/or ampicillin-resistance markers.