New tools for integrated genetic and physical analyses of the Escherichia coli chromosome.
Genetic and biophysical techniques have traditionally been applied to genome mapping independently of one another. We present a series of Escherichia coli mini-Tn10 insertions that contain the rare-cutting polylinker 1 (RCP1) of rare restriction sites [including BlnI/AvrII, SpeI, NheI, XbaI, NotI, PacI and SfiI; Mahillon and Kleckner, Gene 116 (1992) 69-74] which allows them to be used not just for genetic mapping, but also for rapid physical mapping and integrated physical and genetic mapping of the E. coli chromosome. Their isolation and their physical and genetic coordinates in K-12 strain MG1655 are presented. Also, their use in purifying insertion-delimited DNAs from E. coli K-12 and in macrorestriction mapping of a pathogenic strain's chromosome is demonstrated. These insertions allow integration of (i) different macrorestriction patterns of a single strain's chromosome, (ii) the physical map of a single strain's chromosome with the genetic map of the species, and (iii) the physical maps of different strains' chromosomes.
Department of Pediatrics, School of Medicine, University of Michigan, Ann Arbor 48109, USA.
C K
CK
Rode
V H
VH
Obreque
C A
CA
Bloch
D002874:Chromosome Mapping; D002876:Chromosomes, Bacterial / Q000648:ultrastructure*; D004251:DNA Transposable Elements; D004269:DNA, Bacterial; D015252:Deoxyribonucleases, Type II Site-Specific; D004926:Escherichia coli / Q000235:genetics*; D005798:Genes, Bacterial; D015183:Restriction Mapping
D004251:DNA Transposable Elements; D004269:DNA, Bacterial; C064053:CCTAGG-specific type II deoxyribonucleases; D015252:Deoxyribonucleases, Type II Site-Specific; C038565:GCGGCCGC-specific type II deoxyribonucleases