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Infect Immun. Jan 1996; 64(1): 277–282.
PMCID: PMC173756

Characterization of Tn917 insertion mutants of Staphylococcus epidermidis affected in biofilm formation.


Biofilm formation is thought to result from the concerted action of primary attachment to a specific surface and accumulation in multilayered cell clusters. Here we describe the isolation and characterization of transposon (Tn917) mutants of Staphylococcus epidermidis O-47 which were biofilm negative in the polystyrene microtiter plate assay. Among 5,000 Tn917 insertion mutants, 4 biofilm-negative mutants were isolated. Each mutant carried one copy of Tn917. The mutants were divided into two phenotypic classes: class A (mut1 and mut1a) and class B (mut2 and mut2a). Mutants of phenotypic class A lacked four cell surface proteins, were less hydrophobic, and were affected in primary attachment to polystyrene, but were still able to form multilayered cell clusters. They were able to form a biofilm on a glass surface, a trait that was even more pronounced than in the wild-type stain O-47. Loss of several surface proteins might have led to the reduced surface hydrophilic structures, thus favoring primary attachment to a glass surface and leading to subsequent biofilm formation. Mutants of phenotype class B were able to attach to polystyrene but were unable to form multilayered cell clusters, had unchanged cell surface proteins and hydrophobicity, and were unable to form a biofilm on a glass surface, mut1 and mut2 could be complemented by wild-type DNA fragments containing the Tn917 insertion sites of mut1 and mut2, respectively. The complemented biofilm-positive clone mut1 (pRC20) produced a 60-kDa protein which is postulated to function as the adhesin for binding to plastic. The traits of binding to polystyrene and the ability to form multilayered cell clusters are phenotypically and genetically distinct.

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Selected References

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  • Augustin J, Rosenstein R, Wieland B, Schneider U, Schnell N, Engelke G, Entian KD, Götz F. Genetic analysis of epidermin biosynthetic genes and epidermin-negative mutants of Staphylococcus epidermidis. Eur J Biochem. 1992 Mar 15;204(3):1149–1154. [PubMed]
  • Brückner R, Wagner E, Götz F. Characterization of a sucrase gene from Staphylococcus xylosus. J Bacteriol. 1993 Feb;175(3):851–857. [PMC free article] [PubMed]
  • Christensen GD, Barker LP, Mawhinney TP, Baddour LM, Simpson WA. Identification of an antigenic marker of slime production for Staphylococcus epidermidis. Infect Immun. 1990 Sep;58(9):2906–2911. [PMC free article] [PubMed]
  • Christensen GD, Bisno AL, Parisi JT, McLaughlin B, Hester MG, Luther RW. Nosocomial septicemia due to multiply antibiotic-resistant Staphylococcus epidermidis. Ann Intern Med. 1982 Jan;96(1):1–10. [PubMed]
  • Christensen GD, Simpson WA, Bisno AL, Beachey EH. Experimental foreign body infections in mice challenged with slime-producing Staphylococcus epidermidis. Infect Immun. 1983 Apr;40(1):407–410. [PMC free article] [PubMed]
  • Christensen GD, Simpson WA, Younger JJ, Baddour LM, Barrett FF, Melton DM, Beachey EH. Adherence of coagulase-negative staphylococci to plastic tissue culture plates: a quantitative model for the adherence of staphylococci to medical devices. J Clin Microbiol. 1985 Dec;22(6):996–1006. [PMC free article] [PubMed]
  • Franson TR, Sheth NK, Rose HD, Sohnle PG. Scanning electron microscopy of bacteria adherent to intravascular catheters. J Clin Microbiol. 1984 Sep;20(3):500–505. [PMC free article] [PubMed]
  • Hogt AH, Dankert J, Hulstaert CE, Feijen J. Cell surface characteristics of coagulase-negative staphylococci and their adherence to fluorinated poly(ethylenepropylene). Infect Immun. 1986 Jan;51(1):294–301. [PMC free article] [PubMed]
  • Hussain M, Wilcox MH, White PJ. The slime of coagulase-negative staphylococci: biochemistry and relation to adherence. FEMS Microbiol Rev. 1993 Apr;10(3-4):191–207. [PubMed]
  • Johnson GM, Lee DA, Regelmann WE, Gray ED, Peters G, Quie PG. Interference with granulocyte function by Staphylococcus epidermidis slime. Infect Immun. 1986 Oct;54(1):13–20. [PMC free article] [PubMed]
  • Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. [PubMed]
  • Ludwicka A, Jansen B, Wadström T, Pulverer G. Attachment of staphylococci to various synthetic polymers. Zentralbl Bakteriol Mikrobiol Hyg A. 1984 Apr;256(4):479–489. [PubMed]
  • Mack D, Nedelmann M, Krokotsch A, Schwarzkopf A, Heesemann J, Laufs R. Characterization of transposon mutants of biofilm-producing Staphylococcus epidermidis impaired in the accumulative phase of biofilm production: genetic identification of a hexosamine-containing polysaccharide intercellular adhesin. Infect Immun. 1994 Aug;62(8):3244–3253. [PMC free article] [PubMed]
  • Muller E, Hübner J, Gutierrez N, Takeda S, Goldmann DA, Pier GB. Isolation and characterization of transposon mutants of Staphylococcus epidermidis deficient in capsular polysaccharide/adhesin and slime. Infect Immun. 1993 Feb;61(2):551–558. [PMC free article] [PubMed]
  • Novick RP, Bouanchaud D. The problems of drug-resistant pathogenic bacteria. Extrachromosomal nature of drug resistance in Staphylococcus aureus. Ann N Y Acad Sci. 1971 Jun 11;182:279–294. [PubMed]
  • Pascual A, Fleer A, Westerdaal NA, Verhoef J. Modulation of adherence of coagulase-negative staphylococci to Teflon catheters in vitro. Eur J Clin Microbiol. 1986 Oct;5(5):518–522. [PubMed]
  • Patrick CC, Plaunt MR, Hetherington SV, May SM. Role of the Staphylococcus epidermidis slime layer in experimental tunnel tract infections. Infect Immun. 1992 Apr;60(4):1363–1367. [PMC free article] [PubMed]
  • Peters G, Locci R, Pulverer G. Microbial colonization of prosthetic devices. II. Scanning electron microscopy of naturally infected intravenous catheters. Zentralbl Bakteriol Mikrobiol Hyg B. 1981;173(5):293–299. [PubMed]
  • Peters G, Locci R, Pulverer G. Adherence and growth of coagulase-negative staphylococci on surfaces of intravenous catheters. J Infect Dis. 1982 Oct;146(4):479–482. [PubMed]
  • Peters G, Saborowski F, Locci R, Pulverer G. Investigations on staphylococcal infection of transvenous endocardial pacemaker electrodes. Am Heart J. 1984 Aug;108(2):359–365. [PubMed]
  • Pfaller M, Davenport D, Bale M, Barrett M, Koontz F, Massanari RM. Development of the quantitative micro-test for slime production by coagulase-negative staphylococci. Eur J Clin Microbiol Infect Dis. 1988 Feb;7(1):30–33. [PubMed]
  • Pulverer G, Peters G, Schumacher-Perdreau F. Coagulase-negative staphylococci. Zentralbl Bakteriol Mikrobiol Hyg A. 1987 Apr;264(1-2):1–28. [PubMed]
  • Rosenstein R, Peschel A, Wieland B, Götz F. Expression and regulation of the antimonite, arsenite, and arsenate resistance operon of Staphylococcus xylosus plasmid pSX267. J Bacteriol. 1992 Jun;174(11):3676–3683. [PMC free article] [PubMed]
  • Rupp ME, Archer GL. Hemagglutination and adherence to plastic by Staphylococcus epidermidis. Infect Immun. 1992 Oct;60(10):4322–4327. [PMC free article] [PubMed]
  • Schumacher-Perdreau F, Heilmann C, Peters G, Götz F, Pulverer G. Comparative analysis of a biofilm-forming Staphylococcus epidermidis strain and its adhesion-positive, accumulation-negative mutant M7. FEMS Microbiol Lett. 1994 Mar 15;117(1):71–78. [PubMed]
  • Timmerman CP, Fleer A, Besnier JM, De Graaf L, Cremers F, Verhoef J. Characterization of a proteinaceous adhesin of Staphylococcus epidermidis which mediates attachment to polystyrene. Infect Immun. 1991 Nov;59(11):4187–4192. [PMC free article] [PubMed]
  • Tojo M, Yamashita N, Goldmann DA, Pier GB. Isolation and characterization of a capsular polysaccharide adhesin from Staphylococcus epidermidis. J Infect Dis. 1988 Apr;157(4):713–722. [PubMed]

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