Molecular and General Genetics MGG

, Volume 189, Issue 3, pp 382–389 | Cite as

Genetic determinant of pyocin R2 in Pseudomonas aeruginosa PAO

II. Physical characterization of pyocin R2 genes using R-prime plasmids constructed from R68.45
  • Tomoyuki Shinomiya
  • Sawako Shiga
  • Akihiko Kikuchi
  • Makoto Kageyama


The chromosome segment which contains the genes responsible for production of pyocin R2 in P. aeruginosa PAO was defined physically using R-prime plasmids constructed in vivo from R68.45. The previous conclusion from genetic mapping that the cluster of pyocin R2 genes is located in between trpC and trpE genes was confirmed by deletion mapping of various R prime plasmids bearing the trpC gene. The pyocin R2 gene cluster was further localized on two contiguous HinDIII fragments of 16 kb and 8.0 kb. PML14 strain, in which R-type pyocin genes were completely deleted, had only one 11 kb HindIII fragment instead. Heteroduplexes between this 11 kb fragment with the two HindIII fragments of PAO revealed that the cluster of pyocin R2 genes was an insertion 13 kb long.


Pseudomonas Gene Cluster Pseudomonas Aeruginosa Genetic Mapping Chromosome Segment 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Birnboim H, Doly J (1979) A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res 7:1513–1523Google Scholar
  2. Burkardt H-J, Riess G, Pühler A (1979) Relationship of group P1 plasmids revealed by heteroduplex experiments. RP1, RP4, R68 and RK2 are identical. J Gen Microbiol 114:341–348Google Scholar
  3. Daniels D, de Wet J, Blatter F (1980) New map of bacteriophage lambda DNA. J Virol 33:390–400Google Scholar
  4. Davis RW, Botstein D, Roth JR (1980) Advanced bacterial genetics. Cold Spring Harbor Laboratory, New YorkGoogle Scholar
  5. Davis RW, Simon M, Davidson N (1971) Electron microscope heteroduplex methods for mapping regions of base sequence homology in nucleic acids. In: Grossman L, Moldave K (eds) Methods in enzymology, vol 21. Academic Press, New York p 413–428Google Scholar
  6. Denhardt D (1966) A membrane-filter technique for the detection of complementary DNA. Biochem Biophys Res Commun 23:641–652Google Scholar
  7. Haas D, Holloway BW (1976) R factor variants with enhanced sex factor activity in Pseudomonas aeruginosa. Mol Gen Genet 144:243–251Google Scholar
  8. Haas D, Holloway BW, Schamback A, Lesinger T (1977) The genetic organization of arginine biosynthesis in Pseudomonas aeruginosa. Mol Gen Genet 154:7–22Google Scholar
  9. Holloway BW (1969) Genetics of Pseudomonas. Bacteriol Rev 33:419–443Google Scholar
  10. Holloway BW (1978) Isolation and characterization of an R′ plasmid in Pseudomonas aeruginosa. J Bacteriol 133:1078–1082Google Scholar
  11. Ito S, Kageyama M (1970) Relationship between pyocins and a bacteriophage in Pseudomonas aeruginosa. J Gen Appl Microbiol 16:231–240Google Scholar
  12. Ito S, Kageyama M, Egami F (1970) Isolation and characterization of pyocins from several strains of Pseudomonas aeruginosa. J Gen Appl Microbiol 16:205–214Google Scholar
  13. Kageyama M (1975) Bacteriocins and bacteriophages in Pseudomonas aeruginosa. In: Mitsuhashi S, Hashimoto H (eds) Microbial drug resistance. University of Tokyo Press, Tokyo, p 291–305Google Scholar
  14. Kageyama M, Shinomiya T, Aihara Y, Kobayashi M (1979) Characterization of a bacteriophage related to R-type pyocins. J Virol 32:951–957Google Scholar
  15. Krishnapillai V (1971) A novel transducing phage. Its role in recognition of a possible new host-controlled modification system in Pseudomonas aeruginosa. Mol Gen Genet 114:134–143Google Scholar
  16. Leemans J, Villarroel R, Silva B, Van Montagu M, Schell J (1980) Direct repetition of a 1.2 Md DNA sequences is involved in site-specific recombination by the P1 plasmid R68. Gene 10:319–328Google Scholar
  17. Ohsumi M, Shinomiya T, Kageyama M (1980) Comparative study on R-type pyocins of Pseudomonas aeruginosa. J Biochem 87:1119–1126Google Scholar
  18. Riess G, Holloway BW, Pühler A (1980) R68.45, a plasmid with chromosome mobilizing ability (Cma) carries a tandem duplication. Genet Res Camb 36:99–109Google Scholar
  19. Rigby P, Dieckermann M, Rhodes C, Berg P (1977) Labelling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol 113:237–251Google Scholar
  20. Sano Y, Kageyama M (1977) Transformation of Pseudomonas aeruginosa by plasmid DNA. J Gen Appl Microbiol 23:183–186Google Scholar
  21. Shinomiya T (1974) Studies on biosynthesis and morphogenesis of R-type pyocins of Pseudomonas aeruginosa. IV. Quantitative estimation of the rates of synthesis of pyocin R subunit proteins. J Biochem 76:1083–1094Google Scholar
  22. Shinomiya T, Shiga S (1979) Bactericidal activity of the tail of Pseudomonas aeruginosa bacteriophage PS17. J Virol 32:958–967Google Scholar
  23. Shinomiya T, Shiga S, Kageyama M (1983) Genetical determinant of pyocin R2 in Pseudomonas aeruginosa. I. Localization of the cluster of pyocin R2 genes between trpCD and trpE genes. Mol Gen Genet 189:375–381Google Scholar
  24. Southern E (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98:503–517Google Scholar
  25. Willetts N, Crowther C, Holloway BW (1981) The insertion sequence IS21 of R68.45 and the molecular basis for mobilization of the bacterial chromosome. Plasmid 6:30–52Google Scholar
  26. Wu T (1966) A model for three-point analysis of random general transduction. Genetics 54:405–410Google Scholar

Copyright information

© Springer-Verlag 1983

Authors and Affiliations

  • Tomoyuki Shinomiya
    • 1
  • Sawako Shiga
    • 1
  • Akihiko Kikuchi
    • 1
  • Makoto Kageyama
    • 1
  1. 1.Mitsubishi-Kasei Institute of Life SciencesMachida, TokyoJapan

Personalised recommendations