Publication: Development of an intergeneric conjugal transfer system for rimocidin-producing Streptomyces rimosus
Issued Date
2010-01-01
Resource Type
ISSN
1472765X
02668254
02668254
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2-s2.0-77950463542
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Mahidol University
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SCOPUS
Bibliographic Citation
Letters in Applied Microbiology. Vol.50, No.5 (2010), 530-536
Suggested Citation
S. Phornphisutthimas, N. Sudtachat, C. Bunyoo, P. Chotewutmontri, B. Panijpan, A. Thamchaipenet Development of an intergeneric conjugal transfer system for rimocidin-producing Streptomyces rimosus. Letters in Applied Microbiology. Vol.50, No.5 (2010), 530-536. doi:10.1111/j.1472-765X.2010.02835.x Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/29278
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Title
Development of an intergeneric conjugal transfer system for rimocidin-producing Streptomyces rimosus
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Abstract
Aims: To develop an intergeneric conjugation system for rimocidin-producing Streptomyces rimosus. Methods and Results: High efficiencies of conjugation [10-2-10-3transconjugants/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-1MgCl2was 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. © 2010 The Authors. Journal compilation © 2010 The Society for Applied Microbiology.