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Repeated sequences including RS1100 from Pseudomonas cepacia AC1100 function as IS elements.

R A Haugland1, U M Sangodkar, A M Chakrabarty

  • 1Department of Microbiology and Immunology, University of Illinois, College of Medicine, Chicago 60612.

Molecular & General Genetics : MGG
|January 1, 1990
PubMed
Summary

The repeated sequence RS1100 in Pseudomonas cepacia AC1100, now IS931, demonstrates transposition. Another sequence, IS932, was also identified, contributing to genetic mobility and gene expression studies.

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Area of Science:

  • Microbiology
  • Genetics
  • Molecular Biology

Background:

  • Pseudomonas cepacia strain AC1100 possesses a repeated DNA sequence, RS1100.
  • This sequence is located near genes involved in chlorohydroxy hydroquinone (CHQ) degradation.

Purpose of the Study:

  • To investigate the transpositional activity of the RS1100 sequence.
  • To identify and characterize other repeated DNA sequences in P. cepacia AC1100.

Main Methods:

  • DNA hybridization and restriction endonuclease mapping were used to analyze genomic DNA and cosmid clones.
  • Conjugation experiments were performed with recombinant plasmids and a promoter probe plasmid (pKT240).
  • Analysis of reisolated plasmids from recipient strains (P. cepacia AC1100 mutant RHA5 and Escherichia coli transformants) identified sequence insertions.

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Main Results:

  • Evidence confirmed that RS1100 undergoes transposition events.
  • RS1100 copy number and distribution varied among different P. cepacia AC1100 isolates.
  • Transposition of RS1100 into plasmid vectors was observed, sometimes forming tandem copies.
  • Introduction of pKT240 into AC1100 resulted in streptomycin resistance, linked to insertions near the aphC gene.
  • These insertions included RS1100 and a novel 3400 bp repeated sequence.

Conclusions:

  • RS1100 was reclassified as insertion sequence IS931.
  • The 3400 bp repeated sequence was designated as insertion sequence IS932.
  • These findings highlight the dynamic nature of the P. cepacia AC1100 genome and the role of insertion sequences in genetic rearrangements and gene regulation.