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Related Experiment Videos

Characterization of IS199 from Streptococcus mutans V403

F L Macrina1, K R Jones, P Laloi

  • 1Department of Microbiology and Immunology, Virginia Commonwealth University, Richmond 23298-0678, USA. Macrina@GEMS.VCU.EDU

Plasmid
|July 1, 1996
PubMed
Summary

The novel insertion sequence IS199, unique to mutans streptococci, has been sequenced. This element, belonging to the IS3 family, can transpose and its movement can be tracked using an antibiotic resistance gene.

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Genetics

Background:

  • Insertion sequences (IS) are mobile genetic elements found in prokaryotes.
  • Streptococcus mutans is a significant oral bacterium associated with dental caries.
  • Understanding mobile elements in S. mutans is crucial for studying its genome evolution and pathogenicity.

Purpose of the Study:

  • To determine the complete nucleotide sequence of the novel insertion sequence IS199.
  • To characterize IS199's relationship to known insertion sequence families.
  • To investigate the transposition mechanism and frequency of IS199 in Streptococcus mutans.

Main Methods:

  • DNA sequencing to determine the complete nucleotide sequence of IS199.
  • DNA hybridization studies to assess the copy number and distribution of IS199.

Related Experiment Videos

  • Bioinformatic analysis of open reading frames (ORFs) and protein similarities.
  • Transposition assays involving the insertion of an antibiotic resistance gene into IS199.

    • Main Results:

    • IS199 is a 1220 bp insertion sequence found in Streptococcus mutans V403, present in multiple genomic copies.
    • IS199 exhibits characteristics of the IS3 family, including tandem ORFs and sequence similarity in its ORFB product.
    • Transposition of IS199 results in a 3-bp target site duplication, and its movement can be monitored using an engineered antibiotic resistance marker.

    Conclusions:

    • IS199 is a novel member of the IS3 family of insertion sequences, unique to the mutans streptococci.
    • The element appears to transpose efficiently within the S. mutans genome, with potential implications for genomic plasticity.
    • The development of a marked IS199 element facilitates future studies on its transposition dynamics and biological role.