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PMID: 6268941 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Isolation of DNA fragments containing replicating growing forks from both E. coli and B. subtilis.

Molecular & general genetics : MGG ·Vol. 181 ·No. 2 ·1981-00-00 ·Pages 241-7

Valenzuela MS, Aguinaga MD, Inman RB

Abstract

By the use of a restriction enzyme digestion of gently lysed E. coli or B. subtilis cells, it is possible to isolate a minute fraction of the total DNA that has an unusually high sedimentation coefficient. Upon inspection of this DNA in the electron microscope, branched DNA fragments are observed. Single branched DNA fragments were analyzed by restriction enzyme and partial denaturation mapping techniques. The fragments appear to have the properties of growing forks excised from in vivo replicating intermediates. In B. subtilis, the minute fraction of DNA was also investigated by transformation assays and found to be greatly enriched for a marker near the origin and slightly enriched for a terminus marker.

MeSH Terms
Bacillus subtilis/genetics,metabolism Centrifugation, Density Gradient DNA Replication DNA Restriction Enzymes DNA, Bacterial/biosynthesis,isolation & purification Escherichia coli/genetics,metabolism Genetic Markers Microscopy, Electron Nucleic Acid Denaturation
Chemicals
DNA, Bacterial Genetic Markers DNA Restriction Enzymes
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Valenzuela M S
Aguinaga M D
Inman R B
References (16)
16 references, click to expand
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Article Info
Journal
Molecular & general genetics : MGG
Abbr.
Mol Gen Genet
ISSN
0026-8925
DOI
10.1007/BF00268432
Published
1981-00-00
Pages
241-7
Language
English
Region
Germany
NLM ID
0125036
Subset
IM
External Links
PubMed source DOI: 10.1007/BF00268432
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