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IS8- and Tn2353-mediated cointegration of the plasmids R15 and RP4:: Tn1

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Summary

The plasmids R15 and RP4:: Tn1 form fused structures (85 Md and 92 Md cointegrates). The cointegrates do not resolve practically in recA Escherichia coli cells and have a mean life-time of more than 50 generations in a recA + background.

The 85 Md cointegrates were generated at a frequency of 4×10−4 per R15 transconjugant during a mating between E. coli [R15; RP4:: Tn1] and E. coli [F′ColVBtrp:: Tn1755]. These plasmids carry two directly repeated copies of the mobile element IS8 at the junctions between R15 and RP4:: Tn1. The transposition of IS8 from RP4:: Tn1 to the R15 plasmid and the formation of hybrid molecules promoted by this process appear to be induced by the IS8 element of the Tn1755 structure during or after conjugal transfer of F′ColVBtrp:: Tn1755 into E. coli [R15; RP4:: Tn1] cells.

The formation of the 92 Md cointegrates occurs at a frequency of 2×10−5. The fused molecules of R15 and RP4:: Tn1 carry two direct copies of an 8.65 Md R15 fragment at the junctions between these replicons. The fragment has specific features of a new transposon. This element designated Tn2353 determines resistance to Hg, Sm and Su and contains two sites for each BamHI, BglII and SalI and three sites for both EcoRI and PstI. The physical map and some other characteristics of Tn2353 are presented.

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Abbreviations

Ap:

ampicillin

EtBr:

ethidium bromide

Km:

kanamycin

Md:

megadaltons

Sm:

streptomycin

Su:

sulfanilamide

Tc:

tetracycline

[]:

brackets indicate plasmid-carrier state

References

  • Appleyard RK (1954) Segregation of new lysogenic types during growth of a doubly lysogenic strain derived from E. coli K12. Genetics 39:440–452

    Google Scholar 

  • Arthur A, Sherratt DJ (1979) Dissection of the transposition process: A transposon-encoded site-specific recombination system. Mol Gen Genet 175:267–274

    PubMed  Google Scholar 

  • Bachmann BJ (1972) Pedigrees of some mutant strains of Escherichia coli K-12. Bacteriol Rev 36:525–557

    PubMed  Google Scholar 

  • Barth PT, Grinter NJ (1977) Map of plasmid RP4 by insertion of transposon C. J Mol Biol 113:455–474

    PubMed  Google Scholar 

  • Boyer H, Roulland-Dussoix D (1969) A complementation analysis of the restriction and modification of DNA in E. coli. J Mol Biol 41:459–472

    PubMed  Google Scholar 

  • Depicker A, De Block M, Inzé D, Van Montagu M, Shell J (1980) IS-like element IS8 in RP4 plasmid and its involvement in cointegration. Gene 10:329–338

    PubMed  Google Scholar 

  • Dobritsa AP, Dobritsa SV, Tanyashin VI (1978) Isolation and characterization of plasmid from the Bacillus brevis var. G.-B. cells. Mol Gen Genet 164:195–204

    PubMed  Google Scholar 

  • Dobritsa AP, Ksensenko VN, Fedoseeva VB, Alexandrov AA, Kamynina TP, Khmelnitsky MI (1980) Isolation of transposon TnA from plasmid RP4 carying two copies of this element Gene 8:153–162

    PubMed  Google Scholar 

  • Dobritsa AP, Dobritsa SV, Popov EI, Fedoseeva VB (1981) Transposition of a DNA fragment flanked by two inverted Tn1 sequences. Gene 14:217–225

    Article  PubMed  Google Scholar 

  • Dobritsa AP, Ivanova ZA, Fedoseeva VA (1983) Transposition of DNA fragments flanked by two inverted Tn1 sequences: translocation of the plasmid RP4: Tn1 region harboring the Tcr marker. Gene 22:237–243

    Article  PubMed  Google Scholar 

  • Eckhardt T (1978) A rapid method for the identification of plasmid deoxyribonucleic acid in bacteria. Plasmid 1:584–588

    PubMed  Google Scholar 

  • Fredericq P (1969) The recombination of colicinogenic factors with other episomes and plasmids. In: Wolstenholme GEW, O'Connor M (eds) Bacterial episomes and plasmids. Ciba Foundation Symposium, Churchill, London, pp 163–178

    Google Scholar 

  • Galas DJ, Chandler M (1982) Structure and stability of Tn9-mediated cointegrates. Evidence for two pathways of transpositions. J Mol Biol 154:245–272

    PubMed  Google Scholar 

  • Gill RE, Falkow S, Ohtsubo H, Ohtsubo E, So M, Heffron F (1980) A genetic analysis of the transposon Tn3: evidence for cointegrates as intermediates in transposition. In: Mobilization and reassembly of genetic information. Academic Press, London, pp 47–62

    Google Scholar 

  • Gorelov VN, Iljina TS, Smirnov GB (1979) Study of the genetic control of plasmid F′ formation. I. Influence of recA and seg−2 mutations of the donor strain HfrH upon the character of formation of plasmids F′. Genetika (USSR) 15:1206–1220

    Google Scholar 

  • Humpreys GO, Willshaw GA, Anderson ES (1975) A simple method for the preparation of large quantities of pure plasmid DNA. Biochim Biophys Acta 383:457–463

    PubMed  Google Scholar 

  • Meagher RB, Tait RC, Betlach M, Boyer HW (1977) Protein expression in E. coli minicells by recombinant plasmid. Cell 10:521–536

    Article  Google Scholar 

  • Meynell GG, Meynell E (1970) Theory and practice in experimental bacteriology, 2nd ed. Cambridge University Press, Cambridge

    Google Scholar 

  • Miller JH (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York

    Google Scholar 

  • 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

    PubMed  Google Scholar 

  • Tu C-PD, Cohen SN (1980a) Translocation specificity of the Tn3 element: Characterization of sites of multiple insertions. Cell 19:151–160

    Article  PubMed  Google Scholar 

  • Tu C-TD, Cohen SN (1980b) Effect of DNA sequences adjacent to the termini of Tn3 on sequential translocation. Mol Gen Genet 177:597–601

    Article  Google Scholar 

  • Watanabe T, Nishida H, Ogata C, Arai T, Sato S (1964) Episomemediated transfer of drug resistance in Enterobacteriaceae, VII. Two types of naturally occuring R factors. J Bacteriol 88:716–726

    PubMed  Google Scholar 

  • Weston A, Brown MGM, Perkins HR, Saunders JR, Humpreys GO (1981) Transformation of Escherichia coli with plasmid deoxyribonucleic acid: calcium-induced binding of deoxyribonucleic acid to whole cells and to isolated membrane fractions. J Bacteriol 145:780–787

    PubMed  Google Scholar 

  • Willetts NS, Crowther C, Holloway BW (1981) The insertion sequence IS21 of R68.45 and the molecular basis for mobilization of the bacterial chromosome. Plasmid 6:30–52

    PubMed  Google Scholar 

  • Wood WB (1966) Host specificity of DNA produced by E. coli: bacterial mutations affecting the restriction and modification of DNA. J Mol Biol 16:118–133

    PubMed  Google Scholar 

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Communicated by H. Böhme

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Dobritsa, A.P. IS8- and Tn2353-mediated cointegration of the plasmids R15 and RP4:: Tn1 . Molec. Gen. Genet. 194, 206–210 (1984). https://doi.org/10.1007/BF00383518

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