Summary
Two mutants of plasmid RP4 temperaturesensitive for maintenance were isolated and one of them (pTH 10) was extensively studied. Cells carrying pTH 10 showed temperature-sensitive drug resistance from which we isolated a number of temperature-independent derivatives. Almost all of them were Hfrs donating chromosomal genes to recipient bidirectionally from different points of origin. The Hfrs may be formed in two steps: (1) the transposon (Tn 1) carried by pTH 10 translocates into the host chromosome, and (2) pTH 10 is integrated in the host chromosome by reciprocal recombination between the Tn 1 s, one situated on pTH 10 and another on the host chromosome. That temperature-independent drug resistance selects for this type of derivative, was supported by the following observations: (1) Hfrs thus obtained were usually unstable and segregated at high frequency ‘revertants’ showing temperature-sensitive drug resistance when they were cultivated at 30° C. (2) The ‘revertants’, cured of pTH 10 were still ampicillin resistant, indicating existence of Tn 1 inserted in the host chromosome. (3) Tn 1 insertions found in these derivatives mapped in the vicinity of points of origin of the original Hfrs. (4) When new Hfrs were constructed by: (a) transduction with Plkc of Tn 1 insertions found in derivatives of Hfrs, (b) introduction of pTH 10 into the transductants, and (c) isolation of clones of temperature-independent drug resistance from such pTH 10 carrying strains, they had similar characteristics to the original Hfrs from which Tn 1 insertions were derived. Possibilities for genetic manupulation using pTH 10 in a wide range of Gram-negative bacteria are discussed.
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References
Adelberg EA, Mandel M, Chen GCC (1965) Optimal condition for mutagenesis by N-methyl-N′-nitrosoguanidine in Escherichia coli K12. Biochem Biophys Res Commun 18:788–795
Bachmann BJ, Low KB, Taylor AL (1976) Recalibrated linkage map of Escherichia coli K-12. Bacteriol Rev 40:116–167
Barth PT (1979) Plasmid RP4, with Escherichia coli DNA inserted in vitro, mediates chromosomal transfer. Plasmid 2:130–136
Beckwith JR, Signer ER, Epstein W (1966) Transposition of the lac region of E. coli. Cold Spring Harbor Symp Quant Biol 31:393–401
Boucher C, Bergeron B, Barate de Bertalmio M, Dénarié J (1977) Introduction of bacteriophage Mu into Pseudomonas salanacearum and Rhizobium meliloti using the R factor RP4. J Gen Microbiol 98:253–263
Burkardt H-J, Riess G, Pühler A (1979) Relationship of group P1 plasmids revelaed by heteroduplex experiments: RP1, RP4, R68 and PK2 are identical. J Gen Microbiol 114:341–348
Campbell A, Starlinger P, Berg DE, Botstein D, Leaderberg EM, Novick RP, Szybalski W (1979) Nomenclature of transposable elements in prokaryotes. Plasmid 2:466–473
Danilevich VN, Stepanshin YG, Volozhantsev NV, Golub EI (1978) Transposon-mediated insertion of R factor into bacterial chromosome. Mol Gen Genet 161:337–339
Datta N, Hedges RW, Shaw EJ, Sykes RB, Richmond MH (1971) Properties of an R factor from Pseudomonas aeruginosa. J Bacteriol 108:1244–1249
Datta N, Hedges RW (1972) Host ranges of R factors. J Gen Microbiol 70, 453–460
Dénarié J, Rosenberg C, bergeron B, Boucher C, Michel M, Barate de Bertalmio M (1977) Potantial of RP4: Mu plasmids for in vivo genetic engineering of Gram-negative bacteria. In: Bukhari AI, Shapiro JA, Adhya SL (eds) DNA insertion elements, plasmids and episomes, Bold Spring Harbor Press, Cold Spring Harbor, New York, p 507
DiJoseph CG, Bayer ME, Kaji A (1973) Host cell growth in the presence of the thermosensitive drug resistance factor, Rtsl. J Bacteriol 115:399–410
Grinsted J, Bennett PM, Higginson S, Richmond MH (1978) Regional preference of inseriton of Tn501 and Tn802 into RP1 and its derivatives. Mol Gen Genet 166:313–320
Haas D, Holloway BW (1976) R factor variants with enhanced sex factor activity in Pseudomonas aeruginosa. Mol Gen Genet 144:243–251
Harayama S, Palva ET, Hazelbauer GL (1979) Transposon-insertion mutants of Escherichia coli K12 defective in a component common to galactose and ribose chemotaxis. Mol Gen Genet 171:193–203
Holloway BW (1979) Plasmids that mobilize bacterial chromosome. Plasmid 2:1–19
Jacob AE, Cresswell JM, Hedges RW, Coetzee JN, Beringer JE (1976) Properties of plasmids constructed by the in vitro insertion of DNA from Rhizobium leguminsarum or Proteus mirabilis into RP4. Mol Gen Genet 147:315–323
Jaffé-Branchet A, D'Ari R (1977) Maintenance of bacteriophage P1 plasmid. J Viol 23:476–482
Julliot JS, Boistard P (1979) Use of RP4-prime plasmids constructed in vitro to promote a polarized transfer of the chromosome in Escherichia coli and Rhizobium meliloti. Mol Gen Genet 173:289–298
Kingsbury DT, Helinski DR (1973) Temperature-sensitive mutants for the replication of plasmids in Escherichia coli. I. Isolation and specificity of host and plasmid mutations. Genetics 74:17–31
Kleckner N, Roth J, Botstein D (1977) Genetic engineering in vivo using translocatable drug-resistance elements. J Mol Biol 116:125–159
Koyama AH, Yura T (1975) Plasmid mutations affecting selfmaintenance and host growth in Escherichia coli. J Bacteriol 122:80–88
Marrs BL (1979) Conjugative genetics in the photosynthetic bacterium Rhodopseudomonas capsulata. Abstr Ann Meet Am Soc Microbiol H (H) 37 p 142
Martin RR, Sokatch JR, Unger L (1975) Formation of Escherichia coli Hfr strains by integrative suppression with a Pseudomonas R factor. Abstr Ann Meet Am Soc Microbiol H 52 p 104
Meade HM, Signer ER (1977) Genetic mapping of Rhizobium meliloti. Proc Natl Acad Sci USA 74:2076–2078
Miller JH (1972) Experiments in molecular genetics. Cold Spring Harbor Press, Cold Spring Harbor, New York
Nishimura Y, Caro L, Berg CM, Hirota Y (1971) Chromosome replication in Escherichia coli IV. Control of chromosome replication and cell division by an integrated episome. J Mol Biol 55:441–456
Olsen RH, Gonzales C (1974) Escherichia coli gene transfer to unrelated bacteria by a histidine operon-RP 1 drug resistance plamsid complex. Biochem Biophys Res Commun 59:377–385
Olsen RH, Shipley P (1973) Host range and properties of the Pseudomonas aeruginosa R factor R 1822. J Bacteriol 113:772–780
Stanisich VA, Holloway BW (1971) Chromosome transfer in Pseudomonas aeruginosa mediated by R factors. Genet Res Camb 17:169–172
Taylor DE, Levine JG (1979) Characterization of a plasmid mutation affecting maintenance, transfer and elimination by novobiocin. Evidence that bacterial DNA gyrase is required for replication of H group plasmids. Mol Gen Genet 174:127–133
Towner KI (1978) Chromosome mapping in Acinetobacter calcoaceticus. J Gen Microbiol 104:175–180
Towner KJ, Vivian A (1976a) RP4-mediated conjugation in Acinetobacter calcoaceticus. J Gen Microbiol 93:355–360
Towner KJ, Vivian A (1976b) RP4 fertility variants in Acinetobacter calcoaceticus. Genet Res Camb 28:301–306
Towner KJ, Vivian A (1977) Plasmids capable of transfer and chromosome mobilization in Acinetobacter calcoaceticus. J Gen Microbiol 101:167–171
van Vliet F, Silva B, van Montagu M, Schell J (1978) Transfer of RP4: Mu plasmids to Agrobacterium tumefaciens. Plasmid 1:446–455
Watson JM, Holloway BW (1978) Chromosome mapping in Pseudomonas aeruginosa PAT. J Bacteriol 133:1113–1125
Watson MD, Scaife JG (1978) Chromosomal transfer promoted by the promiscuous plasmid RP4. Plasmid 1:226–237
Weinstock GM, Susskind MM, Bostein D (1979) Regional specificity of illegitimate recombination by the translocatable ampicillin-resistance element Tn 1 in the genome of phage P22. Genetics 92:685–710
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Harayama, S., Tsuda, M. & Iino, T. High frequency mobilization of the chromosome of Escherichia coli by a mutant of plasmid RP4 temperature-Sensitive for maintenance. Molec. Gen. Genet. 180, 47–56 (1980). https://doi.org/10.1007/BF00267351
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DOI: https://doi.org/10.1007/BF00267351