High frequency mobilization of gram-negative bacterial replicons by the in vitro constructed Tn5-Mob transposon.
A DNA fragment of the broad host range plasmid RP4 carrying the cis-acting DNA recognition site for conjugative DNA transfer between bacterial cells (Mobsite) was cloned into the kanamycin-neomycin resistance transposon Tn5. Using conventional transposon mutagenesis techniques the new transposon, called Tn5-Mob, can easily be inserted into the host DNA of gram-negative bacteria. A host replicon carrying Tn5-Mob is then mobilizable into any other gram-negative species if the transfer functions of plasmid RP4 are provided in trans. The potential of Tn5-Mob was demonstrated by mobilizing Rhizobium meliloti plasmids as well as the E. coli chromosome at high frequencies.
D002876:Chromosomes, Bacterial / Q000502:physiology; D003001:Cloning, Molecular; D004251:DNA Transposable Elements; D004269:DNA, Bacterial / Q000235:genetics; D004352:Drug Resistance, Microbial; D004926:Escherichia coli / Q000235:genetics; D006090:Gram-Negative Bacteria / Q000235:genetics*; D007612:Kanamycin / Q000494:pharmacology; D009355:Neomycin / Q000494:pharmacology; D011815:R Factors; D012231:Rhizobium / Q000235:genetics
D004251:DNA Transposable Elements; D004269:DNA, Bacterial; D007612:Kanamycin; D009355:Neomycin