Summary
A Pseudomonas TOL plasmid pWW0 possesses toluene degradative pathway (xyl) genes. Unstable maintenance of a pWW0 derivative in Escherichia coli allowed us to identify two transposable elements each carrying all the xyl genes. One element corresponded to a 56 kb transposon, Tn4651, which we had previously characterized. The other element newly identified in this study was 70 kb long, and this element, designated Tn4653, completely included Tn4651. Genetic analysis of Tn4653 demonstrated that its transposition involves two steps, i.e. cointegrate formation and its subsequent resolution. The former step required a trans-acting factor, transposase, which was encoded in a 3.0 kb fragment at one end of Tn4653, and the latter step was inferred to be mediated by the factors necessary for resolution of the Tn4651-mediated cointegrate. The transposase functions were not interchangeable between the two transposons.
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References
Avila P, de la Cruź F, Ward E, Grinsted J (1984) Plasmids containing one inverted repeat of Tn21 can fuse with other plasmids in the presence of Tn21 transposase. Mol Gen Genet 195:288–293
Bagdasarian M, Lurz R, Rückert B, Franklin FCH, Bagdasarian MM, Frey J, Timmis KN (1981) Specific-purpose plasmid clonderived vectors II. Broad host range, high copy number, RSF1010-derived vectors, and a host-vector system for gene cloning in Pseudomonas. Gene 16:237–247
Benson S, Shapiro J (1978) TOL is a broad-host-range plasmid. J Bacteriol 135:278–280
Bolivar F, Rodriguez RL, Greene PJ, Betlach MC, Heyneker HL, Boyer HW, Crosa JH, Falkow S (1977) Construction and characterization of new cloning vehicles II. A multipurpose cloning system. Gene 2:95–113
Clarke L, Carbon J (1976) A colony bank containing synthetic ColE1 hybrid plasmids representative of the entire E. coli genome. Cell 9:91–99
Chang ACY, Cohen SN (1978) Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid. J Bacteriol 134:1141–1156
de la Cruz F, Grinzted J (1982) Genetic and molecular characterization of Tn21, a multiple resistance transposon from R100.1. J Bacteriol 151:222–228
Guyer MS (1983) Uses of the transposon γδ in the analysis of cloned genes. Methods Enzymol 101:362–369
Harayama S, Leppik RA, Rekik M, Mermod N, Lehrbach PR, Reineke W, Timmis KN (1986) Gene order of the TOL catabolic plasmid upper pathway operon and oxidation of both toluene and benzyl alcohol by the xylA product. J Bacteriol 167:455–461
Heffron F (1983) Tn3 and its relatives. In: Shapiro (ed) Mobile genetic elements. Academic Press, New York, pp 223–260
Heritage J, Bennett PM (1985) Plasmid fusions mediated by one end of TnA. J Gen Microbiol 131:1131–1140
Kopecko DJ, Brevet J, Cohen SN (1976) Involvement of multiple translocating DNA segments and recombinational hotspots in the structural evolution of bacterial plasmids. J Mol Biol 108:333–360
Lederberg EM (1987) Plasmid Reference Center Registry of transposon (Tn) and insertion sequence (IS) allocations through December 1986. Gene 51:115–118
Lehrbach PR, Ward J, Meulien P, Broda P (1982) Physical mapping of TOL plasmids pWW0 and pND2 and various R plasmid-TOL derivatives from Pseudomonas spp. J Bacteriol 152:1280–1283
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York
Meulien P, Downing RG, Broda P (1981) Excision of the 40 kb segment of the TOL plasmid from Pseudomonas putida mt-2 involves direct repeats. Mol Gen Genet 184:97–101
Michiels T, Cornelis G (1984) Detection and characterization of Tn2501 a transposon included within the lactose transposon Tn951. J Bacteriol 158:866–871
Mötsch S, Schmitt R (1984) Replicon fusion mediated by a single-ended derivative of transposon Tn1721. Mol Gen Genet 195:281–287
Novick RP, Clowes RC, Cohen SN, Curtiss III R, Datta N, Falkow S (1976) Uniform nomenclature for bacterial plasmids: a proposal. Bacteriol Rev 40:168–189
Peden KWC (1983) Revised sequence of the tetracycline-resistance gene of pBR322. Gene 22:277–280
Simon R, Priefer U, Pühler A (1983) A broad host range mobilization system for in vivo genetic engineering: transposon mutagenesis in Gram negative bacteria. Biotechnology 1:784–791
Timmis KN, Lehrbach PR, Harayama S, Don RH, Mermod N, Bas S, Leppik R, Weightman AJ, Reineke W, Knackmuss HJ (1985) Analysis and manipulation of plasmid-encoded pathways for the catabolism of aromatic compounds by soil bacteria. In: Helinski DR, Cohen SN, Clewell DB, Jackson DA, Hollaender A (eds) Plasmids in bacteria. Plenum Press, New York, pp 719–739
Tsuda M, Iino T (1987) Genetic analysis of a transposon carrying toluene degrading genes on a TOL plasmid pWW0. Mol Gen Genet 210:270–276
Vieira J, Messing J (1982) The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene 19:259–268
Ward JM, Grinsted J (1982) Physical and genetic analysis of the Inc-W group plasmids R388, Sa, and R7K. Plasmid 7:239–250
Williams PA, Murray K (1974) Metabolism of benzoate and the methylbenzoates by Pseudomonas putida (arvilla) mt-2: evidence for the existence of a TOL plasmid. J Bacteriol 120:416–423
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Communicated by M. Takanami
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Tsuda, M., Iino, T. Identification and characterization of Tn4653, a transposon covering the toluene transposon Tn4651 on TOL plasmid pWW0. Molec. Gen. Genet. 213, 72–77 (1988). https://doi.org/10.1007/BF00333400
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DOI: https://doi.org/10.1007/BF00333400