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. 1974 Dec;120(3):1260–1267. doi: 10.1128/jb.120.3.1260-1267.1974

R-Factor Mutant Capable of Specifying Hypersynthesis of Penicillinase

Yoshihiro Odakura 1, Hajime Hashimoto 1, Susumu Mitsuhashi 1
PMCID: PMC245909  PMID: 4612012

Abstract

The physical characteristics of a mutant, RM201-2, capable of conferring high and stable ampicillion resistance was analyzed. The RM201-2 and its parent R-factor deoxyribonucleic acid (DNA) could be isolated as an extrachromosomal and covalently closed circular form. Their buoyant densities were both 1.712 g/cm3, and their molecular weights were about 82 × 106 and 64 × 106, respectively, when measured by CsCl and sucrose density gradient analyses. The contour lengths by electron microscopy were 35.9 ± 0.6 and 31.0 ± 0.6 μm, respectively. By using the extracted R-factor DNA, the mutant and parent characters were transformable to another Escherichia coli strain. The mutant R factor showed an increased amount of DNA even after conjugal transfer to Proteus. An increase in the size of R-factor DNA was thus considered to be the cause of the high level of ampicillin resistance.

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Selected References

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  1. Bruenn J., Hollingsworth H. A mutant of Escherichia coli with a new, highly efficient promoter for the lactose operon. Proc Natl Acad Sci U S A. 1973 Dec;70(12):3693–3697. doi: 10.1073/pnas.70.12.3693. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Clayton D. A., Vinograd J. Circular dimer and catenate forms of mitochondrial DNA in human leukaemic leucocytes. Nature. 1967 Nov 18;216(5116):652–657. doi: 10.1038/216652a0. [DOI] [PubMed] [Google Scholar]
  3. Clowes R. C. Molecular structure of bacterial plasmids. Bacteriol Rev. 1972 Sep;36(3):361–405. doi: 10.1128/br.36.3.361-405.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cohen S. N., Chang A. C., Hsu L. Nonchromosomal antibiotic resistance in bacteria: genetic transformation of Escherichia coli by R-factor DNA. Proc Natl Acad Sci U S A. 1972 Aug;69(8):2110–2114. doi: 10.1073/pnas.69.8.2110. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cohen S. N., Chang A. C. Recircularization and autonomous replication of a sheared R-factor DNA segment in Escherichia coli transformants. Proc Natl Acad Sci U S A. 1973 May;70(5):1293–1297. doi: 10.1073/pnas.70.5.1293. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cohen S. N., Miller C. A. Multiple molecular species of circular R-factor DNA isolated from Escherichia coli. Nature. 1969 Dec 27;224(5226):1273–1277. doi: 10.1038/2241273a0. [DOI] [PubMed] [Google Scholar]
  7. DAVIS B. D., MINGIOLI E. S. Mutants of Escherichia coli requiring methionine or vitamin B12. J Bacteriol. 1950 Jul;60(1):17–28. doi: 10.1128/jb.60.1.17-28.1950. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Davies J. E., Rownd R. Transmissible multiple drug resistance in Enterobacteriaceae. Science. 1972 May 19;176(4036):758–768. doi: 10.1126/science.176.4036.758. [DOI] [PubMed] [Google Scholar]
  9. Goebel W., Schrempf H. Isolation and characterization of supercoiled circular deoxyribonucleic acid from beta-hemolytic strains of Escherichia coli. J Bacteriol. 1971 May;106(2):311–317. doi: 10.1128/jb.106.2.311-317.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hashimoto H., Tanaka T., Mitsuhashi S. Genetic structure of an R factor conferring ampicillin resistance. Jpn J Microbiol. 1973 Sep;17(5):331–337. doi: 10.1111/j.1348-0421.1973.tb00783.x. [DOI] [PubMed] [Google Scholar]
  11. KONDO E., HARADA K., MITSUHASHI S. Drug-resistance of enteric bacteria. 12. Transduction of the transmissible drug resistance factor by bacteriophage Plkc. Jpn J Exp Med. 1962 Feb;32:139–147. [PubMed] [Google Scholar]
  12. Lang D. Molecular weights of coliphages and coliphage DNA. 3. Contour length and molecular weight of DNA from bacteriophages T4, T5 and T7, and from bovine papilloma virus. J Mol Biol. 1970 Dec 28;54(3):557–565. doi: 10.1016/0022-2836(70)90126-9. [DOI] [PubMed] [Google Scholar]
  13. Morris C. F., Hashimoto H., Mickel S., Rownd R. Round of replication mutant of a drug resistance factor. J Bacteriol. 1974 Jun;118(3):855–866. doi: 10.1128/jb.118.3.855-866.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. NOVICK R. P. ANALYSIS BY TRANSDUCTION OF MUTATIONS AFFECTING PENICILLINASE FORMATION IN STAPHYLOCOCCUS AUREUS. J Gen Microbiol. 1963 Oct;33:121–136. doi: 10.1099/00221287-33-1-121. [DOI] [PubMed] [Google Scholar]
  15. Nordström K., Ingram L. C., Lundbäck A. Mutations in R factors of Escherichia coli causing an increased number of R-factor copies per chromosome. J Bacteriol. 1972 May;110(2):562–569. doi: 10.1128/jb.110.2.562-569.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Odakura Y., Tanaka T., Hashimoto H., Mitsuhashi S. Mutation of R factors capable of specifying hypersynthesis of penicillinase. Antimicrob Agents Chemother. 1973 Mar;3(3):315–324. doi: 10.1128/aac.3.3.315. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Odakura Y., Tanaka T., Mitsuhashi S. Drug-resistance and distribution of R factors among Proteus strains. Jpn J Microbiol. 1971 Jul;15(4):367–372. doi: 10.1111/j.1348-0421.1971.tb00593.x. [DOI] [PubMed] [Google Scholar]
  18. Rownd R., Kasamatsu H., Mickel S. The molecular nature and replication of drug resistance factors of the Enterobacteriaceae. Ann N Y Acad Sci. 1971 Jun 11;182:188–206. doi: 10.1111/j.1749-6632.1971.tb30656.x. [DOI] [PubMed] [Google Scholar]
  19. Rownd R., Perlman D., Hashimoto H., Mickel S., Applebaum E., Taylor D. Dissociation and reassociation of the transfer factor and resistance determinants of R factors as a mechanism of gene amplification in bacteria. Johns Hopkins Med J Suppl. 1973;2:115–128. [PubMed] [Google Scholar]
  20. SCHILDKRAUT C. L., MARMUR J., DOTY P. Determination of the base composition of deoxyribonucleic acid from its buoyant density in CsCl. J Mol Biol. 1962 Jun;4:430–443. doi: 10.1016/s0022-2836(62)80100-4. [DOI] [PubMed] [Google Scholar]
  21. Sawai T., Yamagishi S., Mitsuhashi S. Penicillinases of Klebsiella pneumoniae and their phylogenetic relationship to penicillinases mediated by R factors. J Bacteriol. 1973 Sep;115(3):1045–1054. doi: 10.1128/jb.115.3.1045-1054.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Sharp P. A., Cohen S. N., Davidson N. Electron microscope heteroduplex studies of sequence relations among plasmids of Escherichia coli. II. Structure of drug resistance (R) factors and F factors. J Mol Biol. 1973 Apr 5;75(2):235–255. doi: 10.1016/0022-2836(73)90018-1. [DOI] [PubMed] [Google Scholar]

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