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Related Experiment Videos

Spiroplasma plasmids.

S Razin1, I Nur, G Glaser

  • 1Department of Membrane and Ultrastructure Research, Hebrew University-Hadassah Medical School, Jerusalem, Israel.

Israel Journal of Medical Sciences
|June 1, 1987
PubMed
Summary
This summary is machine-generated.

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Researchers identified a novel 8.0-kbp plasmid, pRA1, in Spiroplasma citri. This extrachromosomal DNA can integrate into the spiroplasma chromosome and serves as a DNA probe for detecting S. citri infections.

Area of Science:

  • Microbiology
  • Molecular Biology
  • Genetics

Background:

  • Extrachromosomal DNA, including plasmids, is present in various spiroplasmas, notably Spiroplasma citri.
  • Limited characterization and functional understanding exist for Spiroplasma citri plasmids.

Purpose of the Study:

  • To characterize a novel 8.0-kbp plasmid (pRA1) from Spiroplasma citri (R8A2).
  • To investigate the presence and integration of plasmid sequences in Spiroplasma species.
  • To evaluate pRA1 as a DNA probe for detecting S. citri.

Main Methods:

  • Restriction enzyme mapping and cloning of the pRA1 plasmid.
  • Southern hybridization using nick-translated pRA1 to analyze DNA from different Spiroplasma passage levels and species.
  • Cloning of chromosome-integrated pRA1 sequences into Escherichia coli plasmids.

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Main Results:

  • A distinct 8.0-kbp plasmid, pRA1, was identified and cloned from S. citri (R8A2).
  • Free pRA1 was abundant in low-passage S. citri, while higher passages showed chromosomal integration of plasmid sequences.
  • Integrated plasmid sequences were also found in Spiroplasma kunkelii and Spiroplasma sp. P40.
  • Integrated pRA1 sequences, composed of repetitive elements, did not significantly impact spiroplasma gene expression.
  • Nick-translated pRA1 proved effective as a DNA probe for detecting S. citri in infected plants and insects.

Conclusions:

  • pRA1 represents a novel plasmid in S. citri with a tendency to integrate into the host chromosome.
  • The integration of pRA1 sequences is observed in other Spiroplasma species.
  • pRA1 is a valuable tool for the sensitive detection of Spiroplasma citri.