Summary
Intermolecular transposition of Tn2660 into pCR1 was measured at 30°C in recA − and recA + hosts as between 2.6 and 5.5x10−3, a similar value to that previously found for Tn3. No cointegrate structures were found under conditions where 104 transposition events occurred. Immunity to intermolecular transposition of Tn2660, similar to that found for Tn3 was demonstrated by showing that the above transposition frequency was reduced by a factor of between 10−3 and 10−4 when a mutant Tn2660 (resulting in the synthesis of a temperaturesensitive β-lactamase) was present in the recipient plasmid. Intramolecular transposition of Tn3 was found to occur under the same conditions as previously demonstrated for Tn2660 giving rise to similar end products, in which the newly introduced Tn3 is oriented inversely to the resident Tn3 and the DNA sequence between the two transposons has been inverted. Thus, in all respects functional identity of the transposition activities of Tn3 and Tn2660 is shown, thereby identifying characteristics of intramolecular transposition that are not readily accommodated by current models of transposition.
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References
Arthur A, Sherratt D (1968) Dissection of the transposition process: A transposon-encoded site-specific recombination system. Mol Gen Genet 175: 267–274
Bazaral M, Helinski DR (1968) Circular DNA forms of colicinogenic factors E1, E2 and E3 from Escherichia coli. J Mol Biol 36: 185–194
Chiang SJ, Clowes RC (1980) Intramolecular transposition and inversion in plasmid R6K. J Bacteriol 142: 668–682
Chiang SJ, Clowes RC (1982) Recombination between two TnA transposon sequences oriented as inverse repeats is found less frequently than between direct repeats. Mol Gen Genet 185: 169–175
Clowes RC, Holmans PJ, Chiang SJ (1981) Intramolecular transposition of a β-lactamase sequence and related genetic rearrangements. Cold Spring Harbor Symp Quant Biol 45: 167–171
Covey C, Richardson D, Carbon J (1976) A method for the deletion of restriction sites in bacterial plasmid deoxyribonucleic acid. Mol Gen Genet 145: 155–158
Crosa JH, Luttropp LK, Heffron F, Falkow S (1975) Two replication initiation sites on R-plasmid DNA. Mol Gen Genet 140: 39–50
Crosa JH, Luttropp LK, Falkow S (1976) Mode of replication of the conjugative R-plasmid RSF1040 in Escherichia coli. J Bacteriol 126: 454–466
Dempsey WB, Willetts NS (1976) Plasmid cointegrates of prophage lambda and R factor R100. J Bacteriol 126: 166–176
Gill R, Heffron F, Dougan G, Falkow S (1978) Analysis of sequences transposed by complementation of two classes of transposition-deficient mutants of Tn3. J Bacteriol 136: 742–756
Heffron F, McCarthy BJ, Ohtsubo H, Ohtsubo E (1979) DNA sequence analysis of the transposon Tn3: Three genes and three sites involved in transposition of Tn3. Cell 18: 1153–1163
Heffron F, Rubens C, Falkow S (1975a) Translocation of a plasmid DNA sequence which mediates ampicillin resistance: Molecular nature and specificity of insertion. Proc Natl Acad Sci USA 72: 3623–3627
Heffron F, Sublett R, Hedges R, Jacob A, Falkow S (1975b) Origin of the TEM β-lactamase gene found on plasmids. J Bacteriol 122:p250–256
Hershfield V, Boyer HW, Yanofsky C, Lovett MA, Helinski DR (1974) Plasmid ColE1 as a molecular vehiclefor cloning and amplification of DNA. Proc Natl Acad Sci USA 71: 3455–3459
Inselberg J (1974) Replication of colicin E1 plasmid DNA in minicells from a unique replication initiation site. Proc Natl Acad Sci USA 71: 2256–2259
Kingsbury DT, Helinski DR (1973) Temperature-sensitive mutants for the replication of plasmids in Escherichia coli: Requirement for deoxyribonucleic acid polymerase I in the replication of the plasmid ColE1. J Bacteriol 114: 1116–1124
Kolter R (1981) Replication properties of plasmid R6K. Plasmid 5: 2–9
Kontomichalou P, Mitani M, Clowes RC (1970) Circular R-factor molecules controlling penicillinase synthesis, replicating in Escherichia coli under either relaxed or stringent control. J Bacteriol 104: 34–44
Kopecko DJ, Cohen SN (1975) Site-specific recA-independent recombination between bacterial plasmids: Involvement of palindromes at the recombinational loci. Proc Natl Acad Sci USA 72: 1373–1377
Kostriken R, Morita C, Heffron F (1981) Transposon Tn3 encodes a site-specific recombination system: Identification of essential sequences, genes and actual site of recombination. Proc Natl Acad Sci USA 78: 4041–4045
Kretschmer PJ, Cohen SN (1977) Selected translocation of plasmid genes: Frequency and regional specificity of translocation of the Tn3 element. J Bacteriol 130: 888–899
Lovett MA, Katz L, Helinski DR (1974) Unidirectional replication of plasmid ColE1 DNA. Nature 251: 337–340
Lovett MA, Sparks RB, Helinski DR (1975) Bidirectional replication of plasmid R6K DNA in Escherichia coli: Correspondence between origin of replication and position of single-strand break in relaxed complex. Proc Natl Acad Sci USA 72: 2905–2909
Novick RP, Clowes RC, Cohen SN, Curtiss R III, Datta N, Falkow S (1976) Uniform nomenclature for bacterial plasmids: A proposal. Bacteriol Revs 40: 168–189
Ohtsubo H, Ohmori H, Ohtsubo E (1978) Nucleotide-sequence analysis of Tn3 (A): Implications for insertion and deletion. Cold Spring Harbor Symp Quant Biol 43: 1269–1277
Robinson MK, Bennett PM, Richmond MH (1977) Inhibition of TnA translocation by TnA. J Bacteriol 129: 407–414
Shapiro JA (1979) Molecular model for the transposition and replication of bacteriophage Mu and other transposable elements. Proc Natl Acad Sci USA 76: 1933–1937
So M, Gill R, Falkow S (1975) The generation of a ColE1-Apr cloning vehicle which allows detection of inserted DNA. Mol Gene Genet 142: 239–249
Thorpe PA, Clowes RC (1981) Predicted absence of 5 bp direct repeats flanking transposon sequences. Abstr H66 Ann Gen Mtg ASM, 1981
Tomizawa J-I, Sakakibara Y, Kakefuda T (1974) Replication of colicin E1 plasmid DNA in cell extracts: Origin and direction of replication. Proc Natl Acad Sci USA 71: 2260–2264
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Communicated by G. O'Donovan
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Chiang, SJ., Jordan, E. & Clowes, R.C. Intermolecular and intramolecular transposition and transposition immunity in Tn3 and Tn2660 . Molec Gen Genet 187, 187–194 (1982). https://doi.org/10.1007/BF00331116
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DOI: https://doi.org/10.1007/BF00331116