Abstract
Intracellular location of plasmid NR1 (M = 58 Mg/mol, stringent control of replication, 1–2 copies perEscherichia coli chromosomal equivalent) was studied and compared with that of plasmid R6KΔ1 (M = 21 Mg/mol, relaxed control of replication, 10–15 copies perE. coli chromosomal equivalent), both inE. coli minicells. Considerable difference in relative distribution of molecules of these two plasmid DNA’s between the cytoplasm and the membrane fraction was found when components of the corresponding minicell lyzates were fractionated by sedimentation in a double-linear gradient of caesium chlorid and sucrose. Also the difference in relative numbers of NR1 DNA and R6KΔ1 DNA molecules stably associated with the membrane of minicells, determined by electron-microscopic examination of the fractions containing plasmid DNA-membrane complexes, was evaluated as statistically significant. The association of NR1 DNA molecules withE. coli minicell membrane was found to be a much more frequent event than such association of R6KΔ1 molecules. The absolute amount of plasmid DNA associated with membrane in a single minicell corresponds to one molecule for both NR1 and R6KAΔ1.
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Adler H.I., Fisher W.D., Cohen A., Hardigree A.A.: MiniatureEscherichia coli cells defieient in DNA.Proc.Nat.Acad.Sci. USA 57, 321 (1967).
Clowes R.C.: Molecular structure of bacterial plasmids.Bacteriol.Rev. 36, 361 (1972).
Curtiss J.H.: On transformations used in the analysis of variance.Ann.Math. Statiat. 14, 107 (1943).
Davis R.W., Simon M., Davidson N.: Electron microscope heteroduplex methods for mapping regions of base sequence homology in nucleic acids, p. 413 in L. Grossman, K. Moldave (Eds.):Methods in Enzymology, Vol. XXI. Academic Press, New York —London 1971.
Frazer A.C., Curtiss R. III.: Production, properties and utility of bacteriar minicells.Curr.Top.Microbiol. Immunol. 69, 1 (1975).
Hershfield M.V., LeBlanc D.J., Falkow S.: Membrane association in compatible and incompatibleEscherichia coli host cells following R-factor transfer.J.Bacteriol. 115, 1208 (1973).
Hochmannová J., Nešvera J., Štokbová J.: Visualization of membrane-associated R-plasmid DNA in fraction-ofEscherichia coli minicell lyzate.Folia Microbiol. 25, 11 (1980a).
Hochmannová J., Nešvera J., Štokrová J.: Multicopy plasmid R6KΔ1 inEscherichia coli minicells - interaction of its DNA molecules with the cell membrane, p. 163 in S. Zadražil, J. Šponar (Eds.):DNA: Recombination, Interactions and Repair. Pergamon Press, Oxford 1980a.
Lang D.: Molecular weights of coliphages and coliphage DNA. III. Contour length and molecular weight of DNA from bacteriophage T4, T5 and T7, and from bovine papilloma virus.J. Mol.Biol. 54, 557 (1970).
Mickel S., Arena V. Jr.,Bauer W.: Physical properties and gel electrophoresis behavior of R12-derived plasmid DNAs.Nucleic Acids Res. 4, 1465 (1977).
Nešvera J., Hochmannová J., Štokrová J.: Application of R-plasmid DNA’s fromEscherichia coli minicells in genetic transformation.Folia Microbiol. 23, 278 (1978).
Perlman D., Rownd R.: Transition of R factor NR1 inProteus mirabilis: molecular structure and replication of NR1 deoxyribonucleic acid.J.Bacteriol. 123, 1013 (1975).
Rownd R.H.: Plasmid replication, p. 751 in I. Molineux, M. Kohiyama (Eds.):DNA Synthesis: Present and Future. Plenum, New York 1978.
Rownd R.H., Miki T., Appelbaum E.R., Miller J.R., Finkelstein M., Barton C.R.: Dissociation, amplification, and reassociation of composite R-plasmid DNA, p. 33 in D. Schlessinger (Ed.):Microbiology 1978. Amer. Soc.Microbiol., Washington 1978.
Sparks R.B. Jr.,Helinski D.R.: Association of cellular membrane ofE. coli minicells with the origin/terminus region of replication of plasmid ColEl DNA.Nature 277, 572 (1979).
Sueoka N., Hammers J.: Isolation of DNA-membrane complex inB.subtilis.Proc.Nat.Acod.Sci. USA 71, 4787 (1974).
Worcel A., Burgi E.: Properties of a membrane-attached form of the folded chro mosome ofEecherichia coli.J.Mol.Biol. 82. 91 (1974).
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Hochmannová, J., Nešvera, J., Štokrová, J. et al. Intracellular location of NRl-plasmid DNA inEscherichia coli minicells. Folia Microbiol 26, 270–276 (1981). https://doi.org/10.1007/BF02927250
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DOI: https://doi.org/10.1007/BF02927250