Conjugal transfer of cosmid DNA from Escherichia coli to Saccharopolyspora spinosa: effects of chromosomal insertions on macrolide A83543 production
| Autor: | Jan R. Turner, Patti Matsushima, Mary C. Broughton, Richard H. Baltz |
|---|---|
| Rok vydání: | 1994 |
| Předmět: |
medicine.disease_cause
Insert (molecular biology) chemistry.chemical_compound Plasmid Escherichia coli Genetics medicine Bacteriophages RK2 plasmid Saccharopolyspora spinosa biology General Medicine Cosmids biology.organism_classification Molecular biology Anti-Bacterial Agents Electrophoresis Gel Pulsed-Field chemistry Conjugation Genetic Attachment Sites Microbiological Cosmid Macrolides Homologous recombination DNA Plasmids Saccharopolyspora |
| Zdroj: | Gene. 146:39-45 |
| ISSN: | 0378-1119 |
| DOI: | 10.1016/0378-1119(94)90831-1 |
| Popis: | Cosmid pOJ436, containing large inserts of Saccharopolyspora spinosa (Ss) DNA, was transferred by conjugation from Escherichia coli to Ss an integrated into the chromosome, apparently by homologous recombination, at high frequencies (10(-5) to 10(-4) per recipient). Transfer was mediated by the plasmid RP4 (RK2) transfer functions in E. coli, and the RK2 oriT function located on pOJ436 [Bierman et al., Gene 116 (1992) 43-49]. pOJ436 lacking Ss DNA, or containing a small insert (approx. 2 kb) of Ss DNA, conjugated from E. coli and integrated at either of two bacteriophage phi C31 attB sites at low frequency (approx. 10(-7) per recipient). Exconjugants containing homologous inserts or inserts at the phi C31 attB sites were stable in the absence of antibiotic selection, and most produced control levels of tetracyclic macrolide A83543 factors. Some exconjugants contained similar kinds of large deletions and were defective in macrolide production. |
| Databáze: | OpenAIRE |
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