Publication:
Development of an intergeneric conjugal transfer system for rimocidin-producing Streptomyces rimosus

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Issued Date

2010

Resource Type

Article

File Type

application/pdf

ISSN

2668254

DOI

10.1111/j.1472-765X.2010.02835.x

Other identifier(s)

2-s2.0-77950463542

Rights Holder(s)

Scopus

Bibliographic Citation

Letters in Applied Microbiology. Vol 50, No.5 (2010), p.530-536

Suggested Citation

Phornphisutthimas S., Sudtachat N., Bunyoo C., Chotewutmontri P., Panijpan B., Thamchaipenet A. Development of an intergeneric conjugal transfer system for rimocidin-producing Streptomyces rimosus. Letters in Applied Microbiology. Vol 50, No.5 (2010), p.530-536. doi:10.1111/j.1472-765X.2010.02835.x Retrieved from: https://hdl.handle.net/20.500.14740/7679

Title

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.

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°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. © 2010 The Authors. Journal compilation © 2010 The Society for Applied Microbiology.

Subject(s)

Antibiotics
 Chromosomes
 Escherichia coli
 Fungi
 Molecular biology
 Colony forming units
 Conjugation system
 E. coli
 Gene clusters
 Genetic manipulations
 Green fluorescent protein
 High efficiency
 High stability
 Molecular genetics
 Optimal results
 Southern hybridization
 Streptomyces rimosus
 Streptomycetes
 Transconjugants
 Transfer systems
 Gene expression
 Green fluorescent protein
 Rimocidin
 Polyene
 Rimocidin
 Antibiotics
 Bacterium
 Gene expression
 Hybridization
 Molecular analysis
 Plasmid
 Analytic method
 Article
 Bacterium conjugation
 Bacterium culture
 Colony forming unit
 Controlled study
 DNA sequence
 Gene expression
 Genetic manipulation
 Intergeneric conjugal transfer system
 Mycelium
 Nonhuman
 Nucleotide sequence
 Protein expression
 Sequence analysis
 Southern blotting
 Spore germination
 Streptomyces
 Streptomyces rimosus
 Escherichia coli
 Evaluation study
 Gene transfer
 Genetics
 Metabolism
 Streptomyces
 Actinobacteria (class)
 Streptomyces rimosus
 Streptomycineae
 Conjugation, Genetic
 Escherichia coli
 Gene Transfer Techniques
 Polyenes
 Streptomyces

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URI

https://hdl.handle.net/20.500.14740/7679

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Scopus: Year 1983-2021

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