Summary
The cloacinogenic factor Clo DF13 from Enterobacter cloacae has been transferred to the minicell-producing strain P678-54 of Escherichia coli K12. The data presented show that this Clo DF13 factor segregates into minicells of P678-54 (Clo DF13) and that this factor is the only plasmid, present in these minicells. Proteins from purified P678-54 (Clo DF13) and P678-54 minicells, previously labelled with 14C-amino acids, were compared after electrophoresis on SDS-polyacrylamide gels. From this comparison it appeared that a noninduced Clo DF13 factor directs the synthesis of 4 proteins. The molecular weights of these proteins could be estimated to be about 72000, 32000, 18500 and 12000. In P678-54 (Clo DF13) minicells, one additional Clo DF13 protein was found to be unlabelled. Apparently this protein is not synthesized in P678-54 (Clo DF13) minicells, but is segregated into or is attached to the minicells after being synthesized in the P678-54 (Clo DF13) cells. The molecular weight of this protein is about 62000, which corresponds to the molecular weight of cloacin.
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References
Adler, H. I., Fisher, W. D., Cohen, A., Hardigree, A. A.: Miniature Escherichia coli cells deficient in DNA. Proc. nat. Acad. Sci. (Wash.) 57, 321 (1967).
Bazaral, M., Helinski, D. R.: Circular DNA forms of colicinogenic factors E1, E2 and E3 from Escherichia coli. J. molec. Biol. 36, 185 (1968).
Clayton, D. A., Vinograd, J.: Circular dimer and catenate forms of mitochondrial DNA in human leukaemic leucocytes. Nature (Lond.) 216, 652 (1967).
Cohen, S. N., Silver, R. P., McCoubrey, A. E.: Isolation of catenated forms of R factor DNA from minicells. Nature (Lond.) 231, 249 (1971).
Daniels, M. J., Wild, D. C.: Simple method for preparing polyacrylamide gels for autoradiography. Analyt. Biochem. 35, 544 (1970).
Demerec, M., Adelberg, E. A., Clark, A. J., Hartman, P. E.: A proposal for a uniform nomenclature in bacterial genetics. Genetics 54, 181 (1966).
DeWitt, W., Helinski, D. R.: Characterization of colicinogenic factor E1 from a non-induced and mitomycin C-induced Proteus strain. J. molec. Biol. 13, 692 (1965).
Falkow, S., Haapala, D. K., Silver, R. P., in: Bacterial episomes and plasmids, p. 136. London: Churchill 1969.
Fredericq, P.: Genetics of colicinogenic factors and their relations with other episomes and plasmids. In: L. Ledoux (ed.), Uptake of informative molecules by living cells. Amsterdam: North Holland (in the press).
Godson, G. N., Sinseimer, R. L.: Lysis of Escherichia coli with a neutral detergent. Biochim. biophys. Acta (Amst.) 149, 476 (1967).
Graaf, F. K. de, Planta, R. J., Stouthamer, A. H.: Effect of a bacteriocin produced by Enterobacter cloacae on protein biosynthesis. Biochim. biophys. Acta (Amst.) 240, 122 (1971).
Graaf, F. K. de, Tieze, G. A., Wendelaar Bonga, Sj., Stouthamer, A. H.: Purification and genetic determination of bacteriocin production in Enterobacter cloacae. J. Bact. 95, 631 (1968).
Hudson, B., Vinograd, J.: Catenated circular DNA molecules in HeLa cell mitochondria. Nature (Lond.) 216, 647 (1967).
Inselburg, J.: Segregation into and replication of plasmid deoxyribonucleic acid in chromosomeless segregants of Escherichia coli. J. Bact. 102, 642 (1970).
Inselburg, J., Fuke, M.: Replicating DNA: Structure of colicin factor E1. Science 169, 590 (1970).
Jansz, H., Pouwels, P. S., Schiphorst, J.: Preparation of double-stranded DNA (replicative form) of bacteriophage ϕX174: a simplified method. Biochim. biophys. Acta (Amst.) 123, 626 (1966).
Kass, L. R., Yarmolinski, M. B.: Segregation of functional sex factor into minicells. Proc. nat. Acad. Sci. (Wash.) 66, 815 (1970).
Levy, S. B., Norman, P.: Segregation of transferable R factors into Escherichia coli minicells. Nature (Lond.) 227, 606 (1970).
Novick, R. P.: Extrachromosomal inheritance in bacteria. Bact. Rev. 33, 210 (1969).
Radloff, R., Bauer, W., Vinograd, J.: A dye-buoyant-density method for the detection and isolation of closed circular duplex DNA: The closed circular DNA in HeLa cells. Proc. nat. Acad. Sci. (Wash.) 57, 1514 (1967).
Rhoades, M., MacHattie, L. A., Thomas, C. A., Jr.: The P22 bacteriophage DNA molecule I. The mature form. J. molec. Biol. 37, 21 (1968).
Roozen, K. J., Fenwick, R. G., Jr., Curtiss, R., III: Isolation of plasmids and specific chromosomal segments from Escherichia coli K-12. In: L. Ledoux (ed.), Informative molecules in biological systems, p. 249. Amsterdam: North Holland 1971a.
Roozen, K. J., Fenwick, R. G., Jr., Curtiss, R., III: Synthesis of ribonucleic acid and protein in plasmid-containing minicells of Escherichia coli K-12. J. Bact. 107, 21 (1971b).
Roth, T. F., Hayashi, M.: Allomorphic forms of bacteriophage ϕX174 replicative DNA. Science 154, 658 (1966).
Roth, T. F., Helinski, D. R.: Evidence for circular DNA forms of a bacterial plasmid. Proc. nat. Acad. Sci. (Wash.) 58, 650 (1967).
Stouthamer, A. H., Tieze, G. A.: Bacteriocin production by members of the genus Klebsiella. Antonie v. Leeuwenhoek 32, 171 (1966).
Studier, F. W.: Sedimentation studies of the size and shape of DNA. J. molec. Biol. 11, 373 (1965).
Tieze, G. A., Stouthamer, A. H., Jansz, H. S., Zandberg, J., Bruggen, E. F. J. van: A bacteriocinogenetic factor of Enterobacter cloacae. Molec. gen. Genet. 106, 48 (1969).
Weber, K., Osborn, M.: The reliability of molecular weight determinations by dodecylsulfate polyacrylamide gel electrophoresis. J. biol. Chem. 244, 4406 (1969).
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Kool, A.J., Pranger, M. & Nijkamp, H.J.J. Proteins synthesized by a non-induced bacteriocinogenic factor in minicells of Escherichia coli . Molec. Gen. Genetics 115, 314–323 (1972). https://doi.org/10.1007/BF00333170
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DOI: https://doi.org/10.1007/BF00333170