An improved Tn7-based system for the single-copy insertion of cloned genes into chromosomes of gram-negative bacteria

A system is described for the single-copy, stable insertion of cloned DNA sequences into the chromosomes of Gram − bacteria. Two narrow-host-range plasmids form the basis of this system: the ‘carrier’ plasmid contains the mini Tn7-Km transposon, into which foreign DNA can be cloned; the ‘helper’ plasmid provides the Tn7 transposition functions in trans. Both plasmids are readily transferred into Gram − bacteria by conjugation. The functionality of this system has been demonstrated in Rhodospirillum rubrum.