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Related Experiment Videos

[Plasmid from Halobacterium halobium and its restriction map]

P Shen1, Y Chen

  • 1School of Life Sciences, Wuhan University, Wuchang.

Yi Chuan Xue Bao = Acta Genetica Sinica
|January 1, 1994
PubMed
Summary
This summary is machine-generated.

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Six of ten screened H. halobium strains contain high molecular weight plasmids. The TENS method proved most effective for detection, identifying a specific plasmid pHH205 in strain J7 with 49 copies per cell.

Area of Science:

  • Microbiology
  • Molecular Biology
  • Genetics

Context:

  • Halobacterium halobium is an archaeon studied for its genetic elements.
  • Plasmids are extrachromosomal DNA molecules found in various organisms, including archaea.
  • Understanding plasmid presence and characteristics is crucial for genetic manipulation and research.

Purpose:

  • To screen multiple Halobacterium halobium strains for plasmid DNA.
  • To evaluate different methods for plasmid detection in H. halobium.
  • To characterize a specific plasmid, pHH205, including its form, copy number, molecular weight, and restriction enzyme sites.

Summary:

  • Ten strains of H. halobium were screened for plasmids using four different methods.
  • Six strains were found to harbor high molecular weight plasmids.

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  • The TENS method was identified as the most effective detection technique for H. halobium.
  • Strain J7 contained a single plasmid, pHH205, in a covalently closed circular (CCC) form.
  • Plasmid pHH205 exhibited a molecular weight of 16.7kb and a peak copy number of approximately 49 copies per cell during the late stationary growth phase.
  • Restriction mapping of pHH205 revealed single, adjacent sites for BamHI, HindIII, and XbaI endonucleases.

    • Impact:

    • This study provides valuable insights into the plasmid content of H. halobium strains.
    • The identification of an effective plasmid detection method (TENS) aids future research on H. halobium genetics.
    • Characterization of pHH205, including its copy number and restriction sites, is essential for its use as a potential cloning vector or for genetic studies.