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Development of an intergeneric conjugal transfer system for rimocidin‐producing Streptomyces rimosus
Author(s) -
Phornphisutthimas S.,
Sudtachat N.,
Bunyoo C.,
Chotewutmontri P.,
Panijpan B.,
Thamchaipenet A.
Publication year - 2010
Publication title -
letters in applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.698
H-Index - 110
eISSN - 1472-765X
pISSN - 0266-8254
DOI - 10.1111/j.1472-765x.2010.02835.x
Subject(s) - library science , sociology , computer science
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 MgCl 2 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.