Summary
Tn9 is a transposable element in which a gene (cat) determining chloramphenicol resistance is flanked by directly repeated sequences that are homologous to the insertion sequence IS1. We show here that infection of Escherichia coli K12 (under Rec- Red- Int- conditions) with a λbio transducing phage carrying Tn9 results in the formation of λbio transductants as frequently as cat transductants (about 1 per 106 to 107 infected cells). Most of the λbio transductants do not carry cat, just as most of the cat transductants do not carry λbio. In spite of the absence of cat, the λbio prophage can transpose a second time, from the E. coli chromosome to different sites on an F′ gal plasmid. Analysis of the structure of the transposed λbio element, by restriction nuclease digestion and by electron microscopy, demonstrates that the integrated λbio prophage is flanked by directly repeated IS1 elements. We conclude that there is no genetic information for the ability to transpose encoded in the non-repeated portion of Tn9, i.e. that the directly repeated IS1 elements alone are responsible for Tn9 transposition.
Similar content being viewed by others
References
Berg, D.E.: Genetic evidence for two types of gene arrangements in new λdv plasmid mutants. J. Mol. Biol. 86, 59–68 (1974)
Bukhari, A.I., Froshauer, S.: Isolation of a transposon for chloramphenicol resistance into bacteriophage Mu: Gene 3, 303–314 (1978)
Bukhari, A.I., Shapiro, J.A., Adhya, S.L. (eds.): DNA insertion elements, plasmids, and episomes. New York: Cold Spring Harbor Press 1977
Calos, M., Johnsrud, L., Miller, J.H.: DNA sequence at the integration sites of the insertion elements IS1. Cell 13, 411–418 (1978)
Chandler, M., Roulet, E., Silver, L., Boy de la Tour, E., Caro, L.: Tn10 mediated integration of the plasmid R100.1 into the bacterial chromosome: inverse transposition. Mol. Gen. Genet. 173, 23–30 (1979)
Childs, G.J., Ohtsubo, H., Ohtsubo, E., Sonnenberg, F., Freundlich, M.: Restriction endonuclease mapping of the Escherichia coli K12 chromosome in the vicinity of the ile genes. J. Mol. Biol. 117, 175–193 (1977)
Chow, L.T., Bukhari, A.I.: Heteroduplex electron microscopy of phage Mu mutants containing IS1 insertions and chloramphenicol resistance transposons. Gene 3, 333–346 (1978)
Court, D., Campbell, A.: Gene regulation in N mutants of bacteriophage λ. J. Virol. 9, 938–945 (1972)
DeBruijn, F.J., Bukhari, A.I.: Analysis of transposable elements inserted in the genomes of bacteriophage Mu and P1. Gene 3, 315–331 (1978)
Deonier, R.C., Mirels, L.: Excision of F plasmid sequences by recombination at directly repeated insertion sequence 2 elements: involvement of recA. Proc. Natl. Acad. Sci. U.S.A. 74, 3965–3969 (1977)
Fiandt, M., Hradecna, Z., Lozeron, H.A., Szybalski, W.: Electron micrographic mapping of deletions, insertions, inversions and homologies in the DNAs of coliphage lambda and ph80, pp. 329–354. In: Bacteriophage lambda (A.D. Hershey, ed.) New York: Cold Spring Harbor Press 1971
Fitton, J.E., Packman, L.C., Harford, S., Zaidenzaig, Y., Shaw, W.V.: Plasmids and the evolution of chloramphenicol resistance, pp. 249–252. In: Microbiology-1978 (D. Schlessinger, ed.). Washington, D.C.: ASM Publications 1978
Gill, R., Heffron, F., Dougan, G., Falkow, S.: Analysis of sequences transposed by complementation of two classes of transposition-deficient mutations of Tn3. J. Bacteriol. 136, 742–756 (1978)
Gottesman, M.E., Yarmolinsky, M.B.: Integration-negative mutants of bacteriophage lambda. J. Mol. Biol. 31, 487–505 (1968)
Gottesman, M.M., Rosner, J.L.: Acquisition of a determinant for chloramphenicol resistance by coliphage lambda. Proc. Natl. Acad. Sci. U.S.A. 72, 5041–5045 (1975)
Gottesman, S., Adhya, S.: Genetic, physical, and restriction map of bacteriophage λ, pp. 713–718. In: DNA insertion elements, plasmids, and episomes (A.E. Bukhari, J.A. Shapiro, and S.L. Adhya, eds.). New York: Cold Spring Harbor Press 1977
Grindley, N.D.F.: IS1 insertion generates duplication of a nine base pair sequence at its target site. Cell 13, 419–426 (1978)
Guyer, M.: The λδ sequence of F is an insertion sequence. J. Mol. Biol. 126, 347–365 (1978)
Heffron, F., Bedinger, P., Champoux, J.J., Falkow, S.: Deletions affecting the transposition of an antibiotics resistance gene. Proc. Natl. Acad. Sci. U.S.A. 74, 702–706 (1977)
Johnsrud, L.: DNA sequence of the transposable element IS1. Mol. Gen. Genet. 169, 213–218 (1979)
Johnsrud, L., Calos, M., Miller, J.H.: The transposon Tn9 generates a 9 bp repeated sequence during integration. Cell 15, 1209–1219 (1978)
Kleckner, N.: Translocatable elements in prokaryotes. Cell 11, 11–23 (1978)
Kleekner, N.: DNA sequence analysis of Tn10 insertions: Origin and role of 9 bp flanking repetition during Tn10 translocation. Cell 16, 711–720 (1979)
Kondo, E., Mitsuhashi, S.: Drug resistance of enteric bacteria. IV. Active transducing phage P1CM produced by the combination of R factor with bacteriophage P1. J. Bacteriol. 88, 1266–1274 (1964)
Kondo, E., Mitsuhashi, S.: Drug resistance of enteric bacteria. VI. Introduction of bacteriophage P1CM into Salmonella typhi and formation of P1dCM and F-CM elements. J. Bacteriol. 91, 1787–1794 (1966)
MacHattie, L.A., Jackowski, J.B.: Physical structure and deletion effects of the chloramphenicol resistance element Tn9 in phage lambda, pp. 219–228. In: DNA insertion elements, plasmids, and episomes (Bukhari, A.I., Shapiro, J.A., and Adhya, S.L., eds.). New York: Cold Spring Harbor Press 1977
MacHattie, L.A., Shapiro, J.A.: Chromosomal integration of phage λ by means of a DNA insertion element. Proc. Natl. Acad. Sci. U.S.A. 75, 1490–1494 (1978)
Meyer, R., Boch, G., Shapiro, J.A.: Transposition of DNA inserted into deletions of the Tn5 kanamycin resistance element. Mol. Gen. Genet. 171, 7–13 (1979)
Ohtsubo, E., Hsu, M.T.: Electron microscope heteroduplex studies of sequence relations among plasmids of Escherichia coli: structure of F100, F152, and F8, and mapping of the Escherichia coli chromosomal region pep-supE-gal-attλ-uvrB. J. Bacteriol. 134, 778–794 (1978)
Ohtsubo, E., Lee, H.J., Deonier, R.C., Davidson, N.: Electron microscope heteroduplex relation among plasmids of Escherichia coli. VI. Mapping of F14 sequences homologous to ϕ80dmetBJF and ϕ80dargECBH bacteriophages. J. Mol. Biol. 89, 599–618 (1974)
Ohtsubo, H., Ohtsubo, E.: Nucleotide sequence of an insertion element, IS1. Proc. Natl. Acad. Sci. U.S.A. 75, 615–619 (1978)
Reed, R.R., Young, R.A., Steitz, J.A., Grindley, N.D.F., Guyer, M.S.: Transposition of the E. coli insertion element λδ generates a five base pair repeat. Proc. Natl. Acad. Sci. U.S.A. (in press)
Reif, H.J., Saedler, H.: IS1 is involved in deletion formation in the gal region of E. coli K12. Mol. Gen. Genet. 137, 17–28 (1975)
Robinson, L.H., Landy, A.: HindII, HindIII, and HpaI restriction fragment maps of bacteriophage lambda DNA. Gene 1, 1–31 (1977a)
Robinson, L.H., Landy, A.: HindII, HindIII, and HpaI restriction fragment maps of the left arm of bacteriophage lambda DNA. Gene 1, 33–54 (1977b)
Rosner, J.L.: Formation induction, and curing of bacteriophage P1 lysogens. Virology 48, 679–689 (1972)
Rosner, J.L., Gottesman, M.M.: Transposition and deletion of Tn9: A transposable element carrying the gene for chloramphenicol resistance. pp. 213–218. In: DNA insertion elements, plasmids, and episomes (Bukhari, A.I., Shapiro, J.A., and Adhya, S.L., eds.). New York: Cold Spring Harbor Press 1977
Saedler, H., Heiss, B.: Multiple copies of the insertion DNA sequence IS1 and IS2 in the chromosome of E. coli K12. Mol. Gen. Genet. 122, 267–277 (1973)
Sanger, F., Air, G.M., Barrell, B.G., Brown, N.L., Coulson, A.R., Fidder, J.C., Hutchison, C.A. III, Slocombe, P.M., Smith, M.: Nucleotide sequence of bacteriophage ϕX174 DNA. Nature 265, 687–695 (1977)
Scott, J.R.: Phage P1 cryptic. II. Location and regulation of prophage genes. Virology 53, 327–336 (1973)
Shapiro, J.A.: Molecular model for the transposition and replication of bacteriophage Mu and other transposable elements. Proc. Natl. Acad. Sci. U.S.A. 76, 1933–1937 (1979)
Sharp, P.A., Hsu, M., Ohtsubo, E., Davidson, N.: Electron microscope heteroduplex studies of sequence relations among plasmids of Escherichia coli. I. Structure of F-prime factors. J. Mol. Biol. 71, 471–497 (1972)
Signer, E.R.: Plasmid formation: a new mode of lysogeny by phage λ. Nature 223, 158–160 (1969)
Starlinger, P.: DNA rearrangements in procaryotes. Ann. Rev. Genet. 11, 103–126 (1977)
Starlinger, P., Saedler, H.: IS elements in micro-organisms. Curr. Top. Microbiol. Immunol. 75, 111–152 (1976)
Author information
Authors and Affiliations
Additional information
Communicated by A. Clark
Rights and permissions
About this article
Cite this article
Rosner, J.L., Guyer, M.S. Transposition of IS1-λBIO-IS1 from a bacteriophage λ derivative carrying the IS1-cat-IS1 transposon (Tn9). Molec. Gen. Genet. 178, 111–120 (1980). https://doi.org/10.1007/BF00267219
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00267219