Identification of a promoter sequence in the plasmid pUL340 of Brevibacterium lactofermentum and construction of new cloning vectors for corynebacteria containing two selectable markers

Abstract

A strong promoter P1 has been found in plasmid pUL340, a cloning vector used to transform corynebacteria. This promoter is also expressed efficiently in Escherichia coli. A gene (cat) for chloramphenicol acetyltransferase from Streptomyces acrimycini and a gene (hyg) for hygromycin phosphotransferase from Streptomyces hygroscopicus were subcloned in different positions of the Brevibacterium lactofermentum plasmid pUL340. Both resistance genes are expressed in B. lactofermentum from their own promoters or from the endogenous promoter in pUL340. These genes provide useful screening markers for selecting transformants of B. lactofermentum together with the kanamycin-resistance gene from the transposon Tn5.