Abstract:
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 degrees 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.
journal_name
Lett Appl Microbioljournal_title
Letters in applied microbiologyauthors
Phornphisutthimas S,Sudtachat N,Bunyoo C,Chotewutmontri P,Panijpan B,Thamchaipenet Adoi
10.1111/j.1472-765X.2010.02835.xsubject
Has Abstractpub_date
2010-05-01 00:00:00pages
530-6issue
5eissn
0266-8254issn
1472-765Xpii
LAM2835journal_volume
50pub_type
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