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Infect Immun. Feb 1986; 51(2): 470–475.
PMCID: PMC262355

Molecular alteration of the 140-megadalton plasmid associated with loss of virulence and Congo red binding activity in Shigella flexneri.

Abstract

A plasmid of about 140 megadaltons has been associated with the invasiveness of Shigella flexneri. Upon subculturing in liquid media of fully virulent isolates of Shigella flexneri 2a YSH6000, which contains only a 230-kilobase-pair (kbp) plasmid in addition to 3.3- and 4.2-kbp cryptic plasmids characteristic to all S. flexneri strains, loss of invasiveness, loss of Congo red binding activity (Pcr), and complete loss of, or a deletion, or even a single-site IS insertion in the plasmid occurred simultaneously. This was ascribed to the fact that, once a noninvasive Pcr- cell has emerged, it overgrows the wild type as a consequence of its selective advantage in artificial media. A deletion map of the 230-kbp plasmid was made by analyzing SalI digests of 39 deletion derivatives plus 1 formed by insertion of an IS1-like element in independently isolated, noninvasive Pcr- mutants. Of 39 deletion derivatives, 16 belonged to a single type, and 6 belonged to another, suggesting deletion hot spots. The deletion map was confirmed and extended by analyzing 359 SalI-generated partial digests of the wild-type plasmid cloned into pBR322. Three copies of IS1-like elements were found on three different SalI fragments by Southern hybridization. Segments required for the Pcr+ phenotype seemed to occur at several different locations in the plasmid. Each of 28 representative Pcr- mutants were negative by the Sereny test. Hence, many, or possibly all, Pcr determinants were required for full virulence.

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Selected References

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  • Birnboim HC, Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. [PMC free article] [PubMed]
  • Daskaleros PA, Payne SM. Cloning the gene for Congo red binding in Shigella flexneri. Infect Immun. 1985 Apr;48(1):165–168. [PMC free article] [PubMed]
  • Hale TL, Sansonetti PJ, Schad PA, Austin S, Formal SB. Characterization of virulence plasmids and plasmid-associated outer membrane proteins in Shigella flexneri, Shigella sonnei, and Escherichia coli. Infect Immun. 1983 Apr;40(1):340–350. [PMC free article] [PubMed]
  • Kado CI, Liu ST. Rapid procedure for detection and isolation of large and small plasmids. J Bacteriol. 1981 Mar;145(3):1365–1373. [PMC free article] [PubMed]
  • Labrec EH, Schneider H, Magnani TJ, Formal SB. EPITHELIAL CELL PENETRATION AS AN ESSENTIAL STEP IN THE PATHOGENESIS OF BACILLARY DYSENTERY. J Bacteriol. 1964 Nov;88(5):1503–1518. [PMC free article] [PubMed]
  • Maurelli AT, Blackmon B, Curtiss R., 3rd Loss of pigmentation in Shigella flexneri 2a is correlated with loss of virulence and virulence-associated plasmid. Infect Immun. 1984 Jan;43(1):397–401. [PMC free article] [PubMed]
  • Murayama SY, Sakai T, Makino S, Kurata T, Sasakawa C, Yoshikawa M. The use of mice in the Sereny test as a virulence assay of shigellae and enteroinvasive Escherichia coli. Infect Immun. 1986 Feb;51(2):696–698. [PMC free article] [PubMed]
  • Ohtsubo H, Ohtsubo E. Nucleotide sequence of an insertion element, IS1. Proc Natl Acad Sci U S A. 1978 Feb;75(2):615–619. [PMC free article] [PubMed]
  • Payne SM, Finkelstein RA. Detection and differentiation of iron-responsive avirulent mutants on Congo red agar. Infect Immun. 1977 Oct;18(1):94–98. [PMC free article] [PubMed]
  • Sakai T, Sasakawa C, Makino S, Kamata K, Yoshikawa M. Molecular cloning of a genetic determinant for Congo red binding ability which is essential for the virulence of Shigella flexneri. Infect Immun. 1986 Feb;51(2):476–482. [PMC free article] [PubMed]
  • Sansonetti PJ, Kopecko DJ, Formal SB. Involvement of a plasmid in the invasive ability of Shigella flexneri. Infect Immun. 1982 Mar;35(3):852–860. [PMC free article] [PubMed]
  • Sansonetti PJ, Hale TL, Dammin GJ, Kapfer C, Collins HH, Jr, Formal SB. Alterations in the pathogenicity of Escherichia coli K-12 after transfer of plasmid and chromosomal genes from Shigella flexneri. Infect Immun. 1983 Mar;39(3):1392–1402. [PMC free article] [PubMed]
  • Sasakawa C, Berg DE. IS50-mediated inverse transposition. Discrimination between the two ends of an IS element. J Mol Biol. 1982 Aug 5;159(2):257–271. [PubMed]
  • Sasakawa C, Carle GF, Berg DE. Sequences essential for transposition at the termini of IS50. Proc Natl Acad Sci U S A. 1983 Dec;80(23):7293–7297. [PMC free article] [PubMed]
  • Sasakawa C, Yoshikawa M. Transposon (Tn5)-mediated suppressive integration of ColE1 derivatives into the chromosome of Escherichia coli K12 (dnaA). Biochem Biophys Res Commun. 1980 Oct 16;96(3):1364–1370. [PubMed]
  • SERENY B. Experimental shigella keratoconjunctivitis; a preliminary report. Acta Microbiol Acad Sci Hung. 1955;2(3):293–296. [PubMed]
  • Southern EM. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. [PubMed]
  • Watanabe H, Nakamura A. Large plasmids associated with virulence in Shigella species have a common function necessary for epithelial cell penetration. Infect Immun. 1985 Apr;48(1):260–262. [PMC free article] [PubMed]

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