Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1989 Jul;171(7):3968–3972. doi: 10.1128/jb.171.7.3968-3972.1989

Genetic analysis of Staphylococcus aureus with Tn4001.

G G Mahairas 1, B R Lyon 1, R A Skurray 1, P A Pattee 1
PMCID: PMC210149  PMID: 2544565

Abstract

Tn4001, a 4.5-kilobase composite transposon with IS256 ends that confers resistance to gentamicin (Gmr), tobramycin, and kanamycin in Staphylococcus aureus, can transpose to diverse chromosomal sites in S. aureus. Chromosomal insertions of Tn4001 were isolated either after UV irradiation of transducing lysates carrying pII147::Tn4001 or by selection for thermoresistant Gmr isolates with strains containing thermosensitive derivatives of plasmids pI258 and pII147 carrying Tn4001. Frequent integration of the entire delivery plasmid occurred under these selective conditions in recombination-proficient hosts. When selection for thermoresistant Gmr isolates was done with these plasmids in recombination-deficient hosts, 99% or more of the Gmr isolates resulted from transposition of Tn4001 in the absence of plasmid integration. Efficient isolation of Tn4001 insertions near markers of interest and the isolation of insertional auxotrophs were achieved. Reversion frequencies of insertional auxotrophs were between 10(-6) and 10(-7) (higher than those observed with Tn551 and Tn917). About 50% of the prototrophic revertants were Gms, and these are attributed to precise excision of Tn4001. The Gmr prototrophic revertants were due to intergenic suppression.

Full text

PDF
3970

Images in this article

3971Image
on p.3971

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Barnes I. J., Bondi A., Fuscaldo K. E. Genetic analysis of lysine auxotrophs of Staphylococcus aureus. J Bacteriol. 1971 Feb;105(2):553–555. doi: 10.1128/jb.105.2.553-555.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Goering R. V., Pattee P. A. Mutants of Staphylococcus aureus with increased sensitivity to ultraviolet radiation. J Bacteriol. 1971 Apr;106(1):157–161. doi: 10.1128/jb.106.1.157-161.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Iordanescu S., Surdeanu M. Two restriction and modification systems in Staphylococcus aureus NCTC8325. J Gen Microbiol. 1976 Oct;96(2):277–281. doi: 10.1099/00221287-96-2-277. [DOI] [PubMed] [Google Scholar]
  4. Iordănescu S. Temperature-sensitive mutant of a tetracycline resistance staphylococcal plasmid. Arch Roum Pathol Exp Microbiol. 1976 Jul;35(3):257–264. [PubMed] [Google Scholar]
  5. Jones J. M., Yost S. C., Pattee P. A. Transfer of the conjugal tetracycline resistance transposon Tn916 from Streptococcus faecalis to Staphylococcus aureus and identification of some insertion sites in the staphylococcal chromosome. J Bacteriol. 1987 May;169(5):2121–2131. doi: 10.1128/jb.169.5.2121-2131.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Kleckner N. Transposable elements in prokaryotes. Annu Rev Genet. 1981;15:341–404. doi: 10.1146/annurev.ge.15.120181.002013. [DOI] [PubMed] [Google Scholar]
  7. Luchansky J. B., Pattee P. A. Isolation of transposon Tn551 insertions near chromosomal markers of interest in Staphylococcus aureus. J Bacteriol. 1984 Sep;159(3):894–899. doi: 10.1128/jb.159.3.894-899.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Lyon B. R., Gillespie M. T., Skurray R. A. Detection and characterization of IS256, an insertion sequence in Staphylococcus aureus. J Gen Microbiol. 1987 Nov;133(11):3031–3038. doi: 10.1099/00221287-133-11-3031. [DOI] [PubMed] [Google Scholar]
  9. Lyon B. R., May J. W., Skurray R. A. Tn4001: a gentamicin and kanamycin resistance transposon in Staphylococcus aureus. Mol Gen Genet. 1984;193(3):554–556. doi: 10.1007/BF00382099. [DOI] [PubMed] [Google Scholar]
  10. Lyon B. R., Skurray R. Antimicrobial resistance of Staphylococcus aureus: genetic basis. Microbiol Rev. 1987 Mar;51(1):88–134. doi: 10.1128/mr.51.1.88-134.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. NOVICK R. P., RICHMOND M. H. NATURE AND INTERACTIONS OF THE GENETIC ELEMENTS GOVERNING PENICILLINASE SYNTHESIS IN STAPHYLOCOCCUS AUREUS. J Bacteriol. 1965 Aug;90:467–480. doi: 10.1128/jb.90.2.467-480.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Norrander J., Kempe T., Messing J. Construction of improved M13 vectors using oligodeoxynucleotide-directed mutagenesis. Gene. 1983 Dec;26(1):101–106. doi: 10.1016/0378-1119(83)90040-9. [DOI] [PubMed] [Google Scholar]
  14. Pattee P. A. Chromosomal map location of the alpha-hemolysin structural gene in Staphylococcus aureus NCTC 8325. Infect Immun. 1986 Nov;54(2):593–596. doi: 10.1128/iai.54.2.593-596.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Pattee P. A. Distribution of Tn551 insertion sites responsible for auxotrophy on the Staphylococcus aureus chromosome. J Bacteriol. 1981 Jan;145(1):479–488. doi: 10.1128/jb.145.1.479-488.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Pattee P. A., Neveln D. S. Transformation analysis of three linkage groups in Staphylococcus aureus. J Bacteriol. 1975 Oct;124(1):201–211. doi: 10.1128/jb.124.1.201-211.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Pattee P. A., Thompson N. E., Haubrich D., Novick R. P. Chromosomal map locations of integrated plasmids and related elements in Staphylococcus aureus. Plasmid. 1977 Nov;1(1):38–51. doi: 10.1016/0147-619x(77)90007-5. [DOI] [PubMed] [Google Scholar]
  18. Schroeder C. J., Pattee P. A. Transduction analysis of transposon Tn551 insertions in the trp-thy region of the Staphylococcus aureus chromosome. J Bacteriol. 1984 Feb;157(2):533–537. doi: 10.1128/jb.157.2.533-537.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  20. Stahl M. L., Pattee P. A. Computer-assisted chromosome mapping by protoplast fusion in Staphylococcus aureus. J Bacteriol. 1983 Apr;154(1):395–405. doi: 10.1128/jb.154.1.395-405.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Stahl M. L., Pattee P. A. Confirmation of protoplast fusion-derived linkages in Staphylococcus aureus by transformation with protoplast DNA. J Bacteriol. 1983 Apr;154(1):406–412. doi: 10.1128/jb.154.1.406-412.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Tennent J. M., Lyon B. R., Gillespie M. T., May J. W., Skurray R. A. Cloning and expression of Staphylococcus aureus plasmid-mediated quaternary ammonium resistance in Escherichia coli. Antimicrob Agents Chemother. 1985 Jan;27(1):79–83. doi: 10.1128/aac.27.1.79. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Thompson N. E., Pattee P. A. Transformation in Staphylococcus aureus: role of bacteriophage and incidence of competence among strains. J Bacteriol. 1977 Feb;129(2):778–788. doi: 10.1128/jb.129.2.778-788.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Wyman L., Goering R. V., Novick R. P. Genetic control of chromosomal and plasmid recombination in Staphylococcus aureus. Genetics. 1974 Apr;76(4):681–702. doi: 10.1093/genetics/76.4.681. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Yost S. C., Jones J. M., Pattee P. A. Sequential transposition of Tn916 among Staphylococcus aureus protoplasts. Plasmid. 1988 Jan;19(1):13–20. doi: 10.1016/0147-619x(88)90058-3. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES