Cloning of a plasmid region specifying the N transfer system of bacterial conjugation in Escherichia coli.
HindIII restriction sites were created artificially by the insertion of the transposon Tn5 into the IncN plasmid pCU1 near a presumptive end of its conjugal transfer region (tra). This allowed cloning of an entire and continuous 19.4-kb region of this plasmid that specifies the N transfer system. The cloning vector was the nonconjugative plasmid pACYC184. The recombinant plasmid was as efficient in transfer as the parental N plasmid. Other clones and deletions extending into the tra region allowed localization of a 11.2-kb segment of this region that determines sensitivity to the N-specific bacteriophages IKe and PRD1. It could also be concluded that the ability of pCU1 to promote the killing of Klebsiella pneumoniae requires a 2-kb region that is not part of, but adjacent to the tra region.
D002874:Chromosome Mapping; D003001:Cloning, Molecular; D003227:Conjugation, Genetic; D004251:DNA Transposable Elements; D004269:DNA, Bacterial / Q000235:genetics; D004274:DNA, Recombinant; D004926:Escherichia coli / Q000235:genetics*; D007711:Klebsiella pneumoniae / Q000235:genetics; D010957:Plasmids
D004251:DNA Transposable Elements; D004269:DNA, Bacterial; D004274:DNA, Recombinant