Monitoring of chromosomal insertions of the IncJ elements R391 and R997 in Escherichia coli K-12.
The integration site(s) of the IncJ element, R391, was localised to a specific region of the Escherichia coli chromosome, between the uxuA and serB loci (98.0-99.5 min), using classical Hfr mapping techniques. F-prime plasmid hosts, diploid for regions spanning the E. coli chromosome, were used as recipients in R391 and R997 conjugal transfer assays. Analysis of transconjugants revealed the integration of R391 and R997 into specific F-primes that contain the uxuA to serB region, but not F-primes that contain other regions of the chromosome. A comparison of the electrophoretic mobility of the original F-primes with those containing inserts demonstrated the integration of large elements, in excess of 85 kb. Linear integration of the IncJ elements into chromosomal DNA was demonstrated in recombination-deficient (recA) backgrounds in the absence of detectable autonomous stages. These observations account for the inability to isolate plasmid DNA from IncJ hosts, and suggests that the elements exhibit a conjugative transposon-like biology in E. coli.
Murphy
D B
DB
Department of Chemical and Environmental Science, University of Limerick, Ireland.
Pembroke
J T
JT
D002874:Chromosome Mapping; D002876:Chromosomes, Bacterial / Q000235:genetics*; D003227:Conjugation, Genetic; D004352:Drug Resistance, Microbial / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics* / Q000254:growth & development; D005144:F Factor / Q000235:genetics / Q000302:isolation & purification; D011815:R Factors