| Autor: |
N. Sudtachat, Bhinyo Panijpan, Arinthip Thamchaipenet, Prakitchai Chotewutmontri, Chakrit Bunyoo, Somkiat Phornphisutthimas |
| Rok vydání: |
2010 |
| Předmět: |
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| Zdroj: |
Letters in applied microbiology. 50(5) |
| ISSN: |
1472-765X |
| Popis: |
Aims: To develop an intergeneric conjugation system for rimocidin-producing Streptomyces rimosus. Methods and Results: High efficiencies of conjugation [10−2–10−3 transconjugants/recipient colony forming units (CFU)] were obtained when spores of S. rimosus were heat treated at 40°C for 10 min prior to mixing with E. coli ET12567(pUZ8002/pIJ8600) as donor. Mycelium from liquid grown cultures of S. rimosus could also be used as recipient instead of spores, with 24-h cultures giving optimal results. TSA (Oxoid) medium containing 10 m mol l−1 MgCl2 was the preferred medium for conjugation. Southern hybridization was used to confirm that transconjugants of S. rimosus contained a single copy of pIJ8600 integrated at a unique chromosomal attachment site (attB). The transconjugants exhibited a high stability of plasmid integration and showed strong expression of green fluorescent protein when using pIJ8655 as the conjugative vector. Conclusion: Intergeneric conjugation between E. coli and S. rimosus was achieved at high efficiency using both spores and mycelium. Significance and Impact of the Study: The conjugation system developed provides a convenient gene expression system for S. rimosus R7 and will enable the genetic manipulation of the rimocidin gene cluster. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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