Versatile insertion plasmids for targeted genome manipulations in bacteria: isolation, deletion, and rescue of the pathogenicity island LEE of the Escherichia coli O157:H7 genome

A system of versatile insertion plasmids was constructed that permits efficient delivery of the target sites of an ultra-rare-cutting endonuclease and the recombinase FLP into preselected sites of the bacterial genome. With the help of this system, the pathogenicity island LEE of the Escherichia coli O157:H7 genome was excised and isolated in vitro, deleted in vivo, rescued as a plasmid, and transferred into another strain.

[1]  T. McDaniel,et al.  A cloned pathogenicity island from enteropathogenic Escherichia coli confers the attaching and effacing phenotype on E. coli K‐12 , 1997, Molecular microbiology.

[2]  Purification of Escherichia coli chromosomal segments without cloning. , 1996, Biochemical and biophysical research communications.

[3]  C. Bloch,et al.  New tools for integrated genetic and physical analyses of the Escherichia coli chromosome. , 1995, Gene.

[4]  V. Stewart,et al.  Genetic Analysis of Pathogenic Bacteria: A Laboratory Manual , 1995 .

[5]  T. McDaniel,et al.  A genetic locus of enterocyte effacement conserved among diverse enterobacterial pathogens. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[6]  W. Szybalski,et al.  Escherichia coli genome targeting, I. Cre-lox-mediated in vitro generation of ori- plasmids and their in vivo chromosomal integration and retrieval. , 1994, Gene.

[7]  D. C. Cameron,et al.  Minimizing the Genome of Escherichia coli , 1994 .

[8]  W. Szybalski,et al.  In vivo excision and amplification of large segments of the Escherichia coli genome. , 1994, Nucleic acids research.

[9]  D. C. Cameron,et al.  Minimizing the genome of Escherichia coli. Motivation and strategy. , 1994, Annals of the New York Academy of Sciences.

[10]  W Szybalski,et al.  From the double-helix to novel approaches to the sequencing of large genomes. , 1993, Gene.

[11]  F. Blattner,et al.  Analysis of the Escherichia coli genome: DNA sequence of the region from 84.5 to 86.5 minutes. , 1992, Science.

[12]  M. Ashburner.  A Laboratory manual , 1989 .

[13]  J. Mekalanos,et al.  A novel suicide vector and its use in construction of insertion mutations: osmoregulation of outer membrane proteins and virulence determinants in Vibrio cholerae requires toxR , 1988, Journal of bacteriology.

[14]  D. Helinski,et al.  Binding of purified wild-type and mutant pi initiation proteins to a replication origin region of plasmid R6K. , 1986, Journal of molecular biology.

[15]  Y. Machida,et al.  A 37 X 10(3) molecular weight plasmid-encoded protein is required for replication and copy number control in the plasmid pSC101 and its temperature-sensitive derivative pHS1. , 1984, Journal of molecular biology.

[16]  R. Kolter,et al.  Plasmid R6K DNA replication. I. Complete nucleotide sequence of an autonomously replicating segment. , 1982, Journal of molecular biology.