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Related Experiment Videos

IntI2 integron integrase in Tn7.

Karin Hansson1, Lars Sundström, Alex Pelletier

  • 1Department of Pharmaceutical Biosciences, Division of Microbiology, Uppsala University, Uppsala, Sweden.

Journal of Bacteriology
|March 2, 2002
PubMed
Summary
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This study investigates integrons, mobile genetic elements that facilitate antibiotic resistance gene transfer in bacteria. Researchers characterized the Tn7 integrase (IntI2) and found it shares similarities with the Class 1 integrase (IntI1), revealing insights into antibiotic resistance mobility.

Area of Science:

  • Microbiology
  • Molecular Biology
  • Genetics

Background:

  • Integrons are key elements in bacterial antibiotic resistance, mediating gene acquisition and transfer via site-specific recombination.
  • Class 1 integrons, associated with Tn5090 and Tn21, encode the integrase IntI1.
  • Class 2 integrons are found on transposon Tn7 and related elements, with their integrase gene (intI2) containing a conserved internal stop codon.

Purpose of the Study:

  • To sequence and characterize the Tn7 integrase gene (intI2) and its encoded protein (IntI2*).
  • To investigate the recombination activity of IntI2* and compare it with IntI1.
  • To explore the common pool of mobile antibiotic resistance cassettes utilized by different integron classes.

Main Methods:

  • Sequencing of the intI2 gene from Tn7 and related transposons.

Related Experiment Videos

  • Site-directed mutagenesis to create a functional IntI2* variant (IntI2*179E).
  • Assays to detect site-specific excision of antibiotic resistance cassettes by IntI1 and IntI2*179E.

    • Main Results:

    • The intI2 gene contains a conserved internal stop codon, resulting in a truncated IntI2* peptide (325 amino acids) 46% identical to IntI1.
    • The modified IntI2*179E variant demonstrated site-specific excision of Tn7 cassettes with varying frequencies.
    • Limited cross-specificity was observed; IntI1 excised cassettes from both Tn21 and Tn7, but IntI2*179E did not excise the aadA1 cassette from Tn21.

    Conclusions:

    • Class 1 and Class 2 integrons share a common pool of mobile antibiotic resistance cassettes, facilitating their spread.
    • Integrase IntI1 exhibits broader cassette excision activity than IntI2*179E, suggesting differential roles in resistance gene mobility.
    • The conserved stop codon in intI2 may influence the regulation or function of the Class 2 integrase.