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Related Experiment Videos

Specific single-stranded breaks in mature bacteriophage T7 DNA

S A Khan1, S J Hayes, E T Wright

  • 1Department of Biochemistry, University of Texas Health Science Center, San Antonio 78284-7760, USA.

Virology
|August 1, 1995
PubMed
Summary

Site-specific nicks were found in bacteriophage T7 DNA using denaturing gel electrophoresis. These findings suggest premature T7 terminase activity during DNA packaging is responsible for these specific DNA breaks.

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Area of Science:

  • Molecular Biology
  • Virology
  • Genetics

Background:

  • Bacteriophage T5 DNA exhibits site-specific single-stranded breaks (nicks).
  • Previous methods failed to detect similar nicks in bacteriophage T7 DNA.

Purpose of the Study:

  • To investigate the presence of site-specific nicks in bacteriophage T7 DNA.
  • To characterize the locations and potential causes of these nicks.

Main Methods:

  • Denaturing gel electrophoresis was employed for high-resolution DNA separation.
  • Specific DNA detection using oligonucleotide probes identified nick locations.

Main Results:

  • Site-specific nicks were detected in a fraction of mature bacteriophage T7 DNA molecules.

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  • Nicks were mapped to specific positions on both 5' strands of the T7 genome.
  • The locations of three rightmost nicks correlate with known double-stranded breaks.

    • Conclusions:

    • Bacteriophage T7 DNA, like T5 DNA, possesses site-specific nicks.
    • Premature activation of the T7 terminase during DNA packaging is proposed as the mechanism for nick formation.