Summary
Transposon zdc-235::Tn10 is inserted at min 32 on the genetic map of Escherichia coli, and we have used this transposon to clone 14 kb of DNA that flanks this insertion. The site of insertion of the transposon, and the restriction map of the cloned DNA, correspond well with the predictions of the Bouché restriction map for the terminus region (Bouché 1982). The zdc-235::Tn10 insertion, along with the zdd-230::Tn9 insertion, was used to obtain deletions of the region that has been cloned. Strains lacking a minimum of 14 kb, and more likely a minimum of 40 kb of DNA, showed no alteration of growth or cell morphology.
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Bachmann BJ, Low KB (1980) Linkage map of Escherichia coli K-12, Edition 6. Microbiol Rev 44:1–56
Benton WD, Davis RW (1977) Screening λgt recombinant clones by hybridization to single plaques in situ. Science 196:180–182
Binding R, Romansky G, Bitner R, Kuempel P (1981) Isolation and properties of Tn10 insertions in the rac locus of Escherichia coli. Mol Gen Genet 183:333–340
Bitner RM, Kuempel P (1981) PI transduction map spanning the replication terminus of Escherichia coli K12. Mol Gen Genet 184:208–212
Bitner RM, Kuempel P (1982) P1 transduction mapping of the trg locus in rac + and rac strains of Escherichia coli K12. J Bacteriol 149:529–533
Bochner BR, Huang HC, Schieven GL, Ames BN (1980) Positive selection for loss of tetracycline resistance. J Bacteriol 143:926–933
Bouché JP (1982) Physical map of a 470x103 base-pair region flanking the terminus of DNA replication in the Escherichia coli K12 genome. J Mol Biol 154:1–20
Bouché JP, Gélugne JP, Louarn J, Louarn JM, Kaiser K (1982) Relationships between the physical and genetic maps of a 470x103 base-pair region around the terminus of Escherichia coli K12 DNA replication. J Mol Biol 154:21–32
Clewell DB (1972) Nature of ColE1 plasmid replication in Escherichia coli in the presence of chloramphenicol. J Bacteriol 110:667–676
Clewell DB, Helinski DR (1969) Supercoiled circular DNA-protein complex in Escherichia coli. Proc Natl Acad Sci USA 62:1159–1166
Coleman DC, Foster TJ (1981) Analysis of the reduction in expression of tetracycline resistance determined by Transposon Tn10 in the multicopy state. Mol Gen Genet 182:171–177
Dagert M, Ehrlich SD (1979) Prolonged incubation in calcium chloride improves the competence of Escherichia coli cells. Gene 6:23–28
Daniels DL, Blattner FR (1982) Nucleotide sequence of the Q gene and the Q to S intergenic region of bacteriophage lambda. Virology 117:81–92
Deonier RC, Mirels L (1977) Excision of F plasmid sequences by recombination of directly repeated insertion sequence 2 elements. Proc Natl Acad Sci USA 74:3965–3969
Diderichsen B, Fiil NP, Lavalle R (1977) Genetics of the relB locus in Escherichia coli. J Bacteriol 131:30–33
Enquist L, Sternberg N (1979) In vitro packaging of λDam vectors and their use in cloning DNA fragments. Meth Enz 68:281–301
Fouts KE, Barbour SD (1981) Transductional mapping of ksgB and a new Tn5-induced kasugamycin resistant gene ksgD, in Escherichia coli K12. J Bacteriol 145:914–919
Fouts KE, Barbour SD (1982) Insertions of transposons through the major cotransduction gap of Escherichia coli K12. J Bacteriol 149:106–113
Gottesman M, Gottesman M, Gottesman S, Gellert M (1974) Characterization of bacteriophage λ reverse a an Escherichia coli phage carrying a unique set of host-derived recombination functions. J Mol Biol 88:471–487
Harayama S, Palva ET, Hazelbauer G (1979) Transposon insertion mutants of Escherichia coli K12 defective in a component common to galactose and ribose chemotaxis. Mol Gen Genet 171:193–203
Harayama S, Engstrom P, Wolf-Watz H, Iino T, Hazelbauer GL (1982) Clining of trg, a gene for a sensory transducer in Escherichia coli. J Bacteriol 152:372–383
Hermann RG, Whitfeld PR, Bottomley W (1980) Construction of a SalI/PstI restriction map of spinach chloroplast DNA using low-gelling-temperature agarose electrophoresis. Gene 8:179–181
Horowicz CI, Burke JF (1981) Rapid and efficient cosmid cloning. Nucleic Acids Res 9:2989–2998
Jorgensen RA, Reznikoff WS (1979) Organization of structural and regulatory genes that mediate tetracycline resistance in transposon Tn10. J Bacteriol 138:705–714
Jorgensen RA, Berg DE, Allet B, Reznikoff WS (1979) Restriction enzyme cleavage map of Tn10, a transposon which encodes tetracycline resitance. J Bacteriol 137:681–685
Karn J, Brenner S, Barnett L, Cesareni G (1980) Novel bacteriophage cloning vector. Proc Natl Acad Sci USA 77:5172–5176
Kleckner N, Reichardt K, Botstein D (1979) Inversions and deletions of Salmonella chromosome generated by the translocat-able-tetracycline resistance element Tn10. J Mol Biol 127:89–115
Kuempel P, Duerr S, Seeley N (1977) Terminus region of the chromosome of Escherichia coli inhibits replication forks. Proc Natl Acad Sci USA 74:3927–3931
Kuempel P, Duerr S (1979) Chromosome replication in Escherichia coli is inhibited in the terminus region near the rac locus. Cold Spring Harbor Symp Quant Biol 43:563–567
Lloyd RG, Barbour SD (1974) The genetic location of the sbcA gene of Escherichia coli. Mol Gen Genet 134:157–171
Louarn J, Patte J, Louarn JM (1977) Evidence for a fixed termination site of chromosome replication in Escherichia coli. J Mol Biol 115:295–314
Louarn J, Patte J, Louarn JM (1979) Map position of the replication terminus on the Escherichia coli chromosome. Mol Gen Genet 172:7–11
Low B (1973) Restoration by the rac locus of recombinant forming ability in recB and recC merozygotes of Escherichia coli K12. Mol Gen Genet 122:119–130
Miller JH (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York
Reif HJ, Saedler H (1975) IS1 is involved in deletion formation in the gal region of E. coli K12. J Mol Biol 137:17–28
Rigby PWJ, Dieckmann M, Rhodes C, Berg P (1977) Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol 113:237–251
Ross DG, Grisafi P, Kleckner N, Botstein D (1979a) The ends of Tn10 are not IS3. J Bacteriol 139:1097–1101
Ross DG, Swan J, Kleckner N (1979b) Physical structures, of Tn10-promoted deletions and inversions: role of 1,400 bp inverted repetitions. Cell 16:721–731
Sharp PA, Hsu MT, Ohtsubo E, Davidson N (1972) Electron microscope heteroduplex studies of sequence relations among plasmids of Escherichia coli. I. Structure of F-prime factors. J Mol Biol 71:471–497
Shinnick TM, Lund F, Smithies O, Blattner FR (1975) Hybridization of labeled RNA to DNA in agarose gels. Nucleic Acids Res 2:1911–1929
Skinner MA, Cooper RA (1982) An Escherichia coli mutant defective in the NAD-dependent succinate semialdehyde dehydrogenase. Arch Microbiol 132:270–275
Tsao S, Brunk C, Pearlman R (1983) Protocol for hybridizing dry agarose gels. Analyt Biochem, in press
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Henson, J.M., Kuempel, P.L. The use of transposon insertion zdc-235::Tn10 (min 32) to clone and delete DNA from the terminus region of Escherichia coli . Molec. Gen. Genet. 189, 506–512 (1983). https://doi.org/10.1007/BF00325918
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DOI: https://doi.org/10.1007/BF00325918