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Characterization of a Streptomyces-lividans-type site-specific DNA modification system in the avermectin-producer Streptomyces avermitilis permits investigation of two novel giant linear plasmids, pSA1 and pSA2

Molecular Biology Research Group, School of Biological Sciences, University College of Swansea, Singleton Park, Swansea SA2 8PP, UK
Bioprocess Research, Pfizer Central Research, Groton, CT 06340, USA

^*Author for correspondence: Paul Dyson. Tel: +44 792 295667. Fax: +44 792 295447.

ABSTRACT

The degradation of Streptomyces avermitilis DNA samples analysed by conventional pulsed-field gel electrophoresis was shown to be due to Trisdependent, double-strand cleavage. Using alternative electrophoretic conditions, separation of intact DNA molecules was achieved, permitting the identification of two novel giant linear plasmids: the 100 kb pSA1 and 250 kb pSA2. Use of pSA2 DNA as a probe showed that pSA1 does not cross-hybridize indicating that the plasmids are not closely related. The site-specificity of the DNA modifications, which render the DNA susceptible to Tris-dependent cleavage, was found to be essentially identical to that of similar modifications found in the DNA of S. lividans.

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