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Complete nucleotide sequence of Tn10.

R Chalmers1, S Sewitz, K Lipkow

  • 1Department of Biochemistry, University of Oxford, Oxford OX1 3QU, United Kingdom. chalmers@bioch.ox.ac.uk

Journal of Bacteriology
|April 27, 2000
PubMed
Summary

The complete nucleotide sequence of the Tn10 transposon was determined, revealing a homogeneous genetic unit. It encodes a functional glutamate permease but does not confer heavy metal resistance in E. coli.

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Area of Science:

  • Molecular Biology
  • Genomics
  • Bacterial Genetics

Background:

  • The Tn10 transposon is a well-studied mobile genetic element.
  • Understanding its complete nucleotide sequence is crucial for elucidating its function and evolution.
  • Previous knowledge of Tn10's genetic organization was incomplete.

Purpose of the Study:

  • To determine the complete nucleotide sequence of the Tn10 transposon.
  • To identify novel open reading frames (ORFs) within Tn10.
  • To investigate the functional roles of identified ORFs, including potential roles in transport and resistance.

Main Methods:

  • High-throughput DNA sequencing to determine the complete nucleotide sequence of Tn10.
  • Bioinformatic analysis to identify ORFs and predict protein functions.
  • Gene expression studies to assess the functionality of the glutamate permease.
  • Bacterial growth assays to evaluate heavy metal resistance conferred by Tn10.

Main Results:

  • The complete nucleotide sequence of Tn10 was successfully determined.
  • No discontinuities in dinucleotide signature or G+C content confirmed Tn10's homogeneity.
  • Three novel ORFs were identified: one for a glutamate permease, one for heavy metal resistance repressors, and one for a hypothetical protein in Bacillus subtilis.
  • The glutamate permease was confirmed to be functional upon expression.
  • Tn10 did not provide protection against toxic metal effects in Escherichia coli.

Conclusions:

  • The complete Tn10 sequence reveals a genetically homogeneous element with novel functional ORFs.
  • The identified glutamate permease is functional, suggesting a role in nutrient transport.
  • Tn10 does not confer heavy metal resistance, despite containing ORFs related to resistance operons.
  • Further research is needed to fully understand the role of the hypothetical protein and the repressors in their native contexts.

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