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. 1987 May;169(5):2055–2062. doi: 10.1128/jb.169.5.2055-2062.1987

Use of on-section immunolabeling and cryosubstitution for studies of bacterial DNA distribution.

J A Hobot, M A Bjornsti, E Kellenberger
PMCID: PMC212088  PMID: 3553155

Abstract

Escherichia coli cells were very rapidly frozen and substituted at a low temperature with 3% glutaraldehyde in acetone. Infiltration and embedding with Lowicryl K4M were carried out at -35 degrees C. This procedure resulted in good structural preservation of both the nucleoid morphology and its DNA plasm, such that immunolabeling with the protein-A gold technique could be carried out. With antibodies specific for either double-stranded DNA (dsDNA) or single-stranded DNA (ssDNA), it was shown that dsDNA was present throughout the nucleoid but that ssDNA was located on the nucleoid periphery. Chloramphenicol-treated cells, in which protein synthesis but not DNA replication is stopped, produced a characteristic ringlike nucleoid shape and had both dsDNA and ssDNA present throughout the annular section of the DNA plasm. The relationship between metabolically active DNA and overall bacterial genome organization is discussed.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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