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Molecular and General Genetics MGG

, Volume 217, Issue 2–3, pp 215–222 | Cite as

Genetic deletions between directly repeated sequences in bacteriophage T7

  • James C. Pierce
  • Warren Masker
Article

Summary

DNA sequence analysis of genetic deletions in bacteriophage T7 has shown that these chromosomal rearrangements frequently occur between directly repeated DNA sequences. To study this type of spontaneous deletion in more quantitative detail synthetic fragments of DNA, made by hybridizing two complementary oligonucleotides, were introduced into the non-essential T7 gene 1.3 which codes for T7 DNA ligase. This insert blocked synthesis of functional ligase and made the phage that carried an insert unable to form plaques on a host strain deficient in bacterial ligase. The sequence of the insert was designed so that after it is put into the T7 genome the insert is bracketed by direct repeats. Perfect deletion of the insert between the directly repeated sequences results in a wild-type phage. It was found that these deletion events are highly sensitive to the length of the direct repeats at their ends. In the case of 5 bp direct repeats excision from the genome occurred at a frequency of less than 10−10, while this value for an almost identical insert bracketed by 10 bp direct repeats was approximately 10−6. The deletion events were independent of a hostrecA mutation.

Key words

Deletions Bacteriophage T7 Direct repeats 

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Copyright information

© Springer-Verlag 1989

Authors and Affiliations

  • James C. Pierce
    • 1
  • Warren Masker
    • 1
  1. 1.Department of BiochemistryTemple University School of MedicinePhiladelphiaUSA

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