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Direct repeats flanking the Bacteroides transposon Tn4351 are insertion sequence elements.

V Hwa1, N B Shoemaker, A A Salyers

  • 1Department of Microbiology, University of Illinois, Urbana 61801.

Journal of Bacteriology
|January 1, 1988
PubMed
Summary

The direct repeats flanking the clindamycin-erythromycin resistance region of Bacteroides transposon Tn4351 are insertion sequence (IS) elements. IS4351L mediated chloramphenicol marker transfer more efficiently than IS4351R, revealing differential IS-mediated activities.

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Genetics

Background:

  • The Bacteroides transposon Tn4351 contains a clindamycin-erythromycin resistance (Ccr Emr) region.
  • This resistance region is flanked by direct repeat sequences.

Purpose of the Study:

  • To identify the nature of the direct repeat sequences flanking the Ccr Emr region of Tn4351.
  • To investigate the role of these direct repeats in mediating the transfer of antibiotic resistance markers.

Main Methods:

  • Characterization of flanking direct repeats as insertion sequence (IS) elements.
  • Conjugation experiments using a chloramphenicol (Cmr) marker on pBR328 and the IncW plasmid R388.
  • Analysis of plasmids from transconjugants to identify IS-mediated events.

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Main Results:

  • The direct repeats flanking the Ccr Emr region were identified as IS elements (IS4351L and IS4351R).
  • Both IS elements mediated transfer of the Cmr marker via cointegrate formation with R388.
  • IS4351L-mediated transfer occurred at a higher frequency compared to IS4351R-mediated transfer.
  • Analysis revealed IS-mediated deletions, tandem duplication of IS4351L, and excision of IS4351R.

Conclusions:

  • The direct repeats flanking the Ccr Emr region of Tn4351 are functional IS elements.
  • Differential frequencies of marker transfer mediated by IS4351L and IS4351R were observed.
  • These IS elements exhibit various DNA manipulation activities, including deletion, duplication, and excision.