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PMID: 8981988 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Denaturation of circular or linear DNA facilitates targeted integrative transformation of Streptomyces coelicolor A3(2): possible relevance to other organisms.

Journal of bacteriology ·Vol. 179 ·No. 1 ·1997-01-00 ·Pages 122-7

Oh SH, Chater KF

Abstract

Using Streptomyces coelicolor A3(2) protoplasts, the number of transformants obtained by homologous recombination of incoming double-stranded circular DNA with the recipient chromosome was greatly stimulated by simple denaturation of the donor DNA. This procedure was very effective with inserts over a ca. 100-fold size range, the largest tested being ca. 40-kb inserts in cosmids. These observations led to transformation experiments with linearized cloned DNA and randomly sheared genomic DNA. In both cases, DNA denaturation led to significant levels of transformation. Most of the transformants had resulted from the predicted homologous recombination events. A number of genetic manipulations will be made easier or possible by these procedures.

MeSH Terms
Bacterial Proteins/genetics DNA Methylation DNA, Bacterial/chemistry DNA, Circular/chemistry Genes, Bacterial/genetics Molecular Weight Nucleic Acid Denaturation Plasmids/chemistry Streptomyces/genetics Transcription Factors/genetics Transformation, Bacterial
Chemicals
Bacterial Proteins DNA, Bacterial DNA, Circular Transcription Factors whiB protein, Streptomyces
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Oh S H
Department of Genetics, John Innes Centre, Colney, Norwich, United Kingdom.
Chater K F
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.179.1.122-127.1997
Published
1997-01-00
Pages
122-7
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC178669
Subset
IM
External Links
PubMed source DOI: 10.1128/jb.179.1.122-127.1997 PMC Full Text View journal details
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