A gene cloning system for the siomycin producer <Emphasis Type="Italic">Streptomyces sioyaensis</Emphasis> NRRL-B5408 |
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Authors: | M Myronovskyy B Ostash I Ostash V Fedorenko |
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Institution: | (1) Department of Genetics and Biotechnology of Ivan Franko National University of Ľviv, Ľviv, Ukraine |
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Abstract: | Streptomyces sioyaensis NRRL-B5408 produces a siomycin complex (a group of thiopeptide antibiotics structurally related to thiostrepton). Development
of genetic tools for the detection of siomycin production and DNA transfer into this strain is described. The existing tipA-based reporter system for determination of siomycin production was modified to achieve its stable integration into actinomycete
genomes. Various replicative plasmids (pKC1139, pKC1218E, pSOK101) as well as actinophage ϕC31- and VWB-based vectors pSET152
and pSOK804, respectively, were conjugally transferred from E. coli into the siomycin producer at a frequency ranging from 3.7 × 10−9 to 1.1 × 10−5. The transconjugants did not differ from wild type in their ability to produce siomycin. There is one attB site for each integrative plasmid. The utility of temperature sensitive replicon of pKC1139 for insertional gene inactivation
in S. sioyaensis has been validated by disruption of putative nonribosomal peptide synthetase gene. |
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