Courtright, James B.

Rohatgi, Kalpana

Webb, Gene

English

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Marquette Universitye-Publications@MarquetteBiological Sciences Faculty Research andPublications Biological Sciences, Department of12-1-1990Location of gyrA on the Physical Map of theEscherichia coli ChromosomeGene WebbMarquette UniversityKalpana RohatgiMarquette UniversityJames B. CourtrightMarquette University, james.courtright@marquette.eduPublished version. Journal of Bacteriology, Vol. 172, No. 12 (December 1990): 6617. Permalink. ©1990 American Society for Microbiology. Used with permission.JOURNAL OF BACTERIOLOGY, Dec. 1990, p. 66170021-9193/90/126617-01$02.00/0Copyright © 1990, American Society for MicrobiologyVol. 172, No. 12Location of gyrA on the Physical Map of theEscherichia coli ChromosomeGENE WEBB, KALPANA ROHATGI, AND JAMES B. COURTRIGHT*Department ofBiology, Marquette University, Milwaukee, Wisconsin 53233Three oligonucleotide probes with sequences correspond-ing to either the coding sequence or the complementarysequence of gyrA (6, 7) at nucleotide positions 295, 343, and524 were synthesized. These oligomers were separatelylabeled at their 5' ends with 32P and hybridized to phageDNA prepared from the X miniset phages 371 through 377(4). Each of the three probes hybridized only to E13A5(phage 376). pAW012, which contains an EcoRV genomicfragment containing the gyrA gene (7), hybridized to 4F12(phage 375) as well as to E13A5. The pAW012 BamHI-HindIII fragment, which contains only the first 692 nucleo-tides for the gyrA gene (6, 7), did-not hybridize to 4F12. ThegyrA gene contains restriction sites for HindIII, KpnI, PstI,and PvuII but none for the other four enzymes used fordefining sites in the recombinant phage collection (4). Thiscombined information identifies the HindIII site at nucleo-tide 692 of gyrA with the leftmost HindIII site of the E13A5insert. The fact that the oligonucleotides hybridized with thisphage, whereas pAW012 also hybridized to phage strain4F12, places the gyrA gene, map position 48.3 min (1), atcoordinate 2350 (5) immediately to the left of ubiG and withan orientation the same as that of ubiG (2) but opposite thatof the atoDAB (3) operon at coordinate 2334 (Fig. 1). Therewas no inconsistency in terms of restriction sites, and thesizes of cloned genes were in good agreement with therestriction map (4, 5).* Corresponding author.2330 2340 2350HH B2360RL-0 WIatoDAB gyrA ubiG nrdABFIG. 1. Organization of the region encompassing coordinates2330 to 2350 of the Escherichia coli chromosome. The coordinatesof atoDAB, gyrA, ubiG, and nrdAB at BamHI (B), EcoRI (R), andHindIII (H) sites are 2336.5, 2350, 2352.6, and 2357.68, respectively,on the basis of the corrected restriction map (5).ACKNOWLEDGMENTSThis work was supported by Biomedical Research Support Grantto Marquette University and the Parke-Davis Pharmaceutical Re-search Division.LITERATURE CITED1. Bachmann, B. J. 1990. Linkage map of Escherichia coli K-12,edition 8. Microbiol. Rev. 54:130-197.2. Gfbert, I., M. Llagostera, and J. Barbe. 1988. Regulation of ubiGexpression in Escherichia coli. J. Bacteriol. 170:1346-1349.3. Jenkins, L. S., and W. D. Nunn. 1987. Genetic and molecularcharacterization of the genes involved in short-chain fatty aciddegradation in Escherichia coli: the ato system. J. Bacteriol. 169:42-52.4. Kohara, Y., K. Akiyama, and K. Isono. 1987. The physical map ofthe whole E. coli chromosome: application of a new strategy forrapid analysis and sorting of a large genomic library. Cell 50:495-508.5. MWdigue, C., J. P. Bouch6, A. Hinart, and A. Danchin. 1990.Mapping of sequenced genes (700kbp) in the restriction map ofthe Escherichia coli chromosome. Mol. Microbiol. 4:169-187.6. Swanberg, S. L., and J. C. Wang. 1987. Cloning and sequencihgof the Escherichia coli gyrA gene coding for the A subunit ofDNA gyrase. J. Mol. Biol. 197:729-736.7. Yoshida, H., Y. Kojima, J. Yamagishi, and S. Nakamura. 1988.Quinolone-resistant mutations of the gyrA gene of Escherichiacoli. Mol. Gen. Genet. 211:1-7.6617

Location of \u3cem\u3egyrA\u3c/em\u3e on the Physical Map of the \u3cem\u3eEscherichia coli\u3c/em\u3e Chromosome

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Location of \u3cem\u3egyrA\u3c/em\u3e on the Physical Map of the \u3cem\u3eEscherichia coli\u3c/em\u3e Chromosome

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