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Gene. 1991 Dec 20;109(1):167-8.

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

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Department of Bacteriology, College of Agricultural and Life Sciences, University of Wisconsin, Madison 53706.


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.

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