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Related Experiment Videos

ISRf1, a transposable insertion sequence from Sinorhizobium fredii

J M Vinardell1, F J Ollero, H B Krishnan

  • 1Departamento de Microbiología, Facultad de Biología , Universidad de Sevilla, Spain.

Gene
|January 20, 1998
PubMed
Summary
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A novel insertion sequence, ISRf1, found in Sinorhizobium fredii HH103, can activate a silent tetracycline resistance gene. This mobile genetic element has implications for understanding gene regulation in nitrogen-fixing bacteria.

Area of Science:

  • Microbiology
  • Molecular Biology
  • Genetics

Background:

  • Sinorhizobium fredii HH103 is a nitrogen-fixing bacterial symbiont.
  • Mobile genetic elements, like insertion sequences (IS), play roles in genome evolution and gene expression.
  • Plasmid pMUS248 in HH103 contains a promoterless tetracycline resistance (TcR) gene.

Purpose of the Study:

  • To clone and characterize a novel insertion sequence (IS) element in Sinorhizobium fredii HH103.
  • To investigate the mechanism by which this IS element activates the promoterless TcR gene on plasmid pMUS248.

Main Methods:

  • Cloning and sequencing of the insertion sequence element.
  • Characterization of the element's structural features, including length, open reading frame (ORF), and terminal inverted repeats.

Related Experiment Videos

  • Analysis of the IS element's presence in other bacterial strains and its transposition frequency.

    • Main Results:

    • The insertion sequence element, designated ISRf1, is 1002 bp long with a 513-bp ORF and 36-bp terminal inverted repeats.
    • ISRf1 creates 5-bp target duplications upon insertion.
    • Two copies of ISRf1 are present in the HH103 genome but absent in 12 other Sinorhizobium and Rhizobium strains.
    • ISRf1 exhibits a transposition frequency of 2.7 x 10(-6) per generation per cell.

    Conclusions:

    • ISRf1 is a novel, transposable insertion sequence in Sinorhizobium fredii HH103.
    • ISRf1 has the capacity to activate a promoterless TcR gene, suggesting a role in horizontal gene transfer and adaptation.
    • The specific distribution of ISRf1 indicates it is likely a recent acquisition or has limited mobility across diverse strains.