Denaturation of circular or linear DNA facilitates targeted integrative transformation of Streptomyces coelicolor A3(2): possible relevance to other organisms.
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.
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S H
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Department of Genetics, John Innes Centre, Colney, Norwich, United Kingdom.
Chater
K F
KF
D001426:Bacterial Proteins / Q000235:genetics; D019175:DNA Methylation; D004269:DNA, Bacterial / Q000737:chemistry*; D004270:DNA, Circular / Q000737:chemistry*; D005798:Genes, Bacterial / Q000235:genetics; D008970:Molecular Weight; D009691:Nucleic Acid Denaturation; D010957:Plasmids / Q000737:chemistry; D013302:Streptomyces / Q000235:genetics*; D014157:Transcription Factors / Q000235:genetics; D014169:Transformation, Bacterial
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D004270:DNA, Circular; D014157:Transcription Factors; C081543:whiB protein, Streptomyces