Mutation and cloning of clustered Streptomyces genes essential for sulphate metabolism |
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Authors: | Derek J. Lydiate Carmen Mendez Helen M. Kieser David A. Hopwood |
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Affiliation: | (1) John Innes Institute, Colney Lane, NR4 7UH Norwich, UK;(2) Present address: Max-Planck-Institut für Züchtungsforschung, D-5000 Köln 30, Germany;(3) Present address: Departamento Interfacultativo de Microbiologia, Universidad de Oviedo, Oviedo, Spain |
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Abstract: | Summary A range of mutants auxotrophic for cysteine (cys) and resistant to selenate (sel) were isolated from many Streptomyces strains but chiefly from S. coelicolor A3(2) and S. lividans 66. Two of the classes of sel/cys mutants probably contained simple biochemical lesions of sulphate permease (selC) and ATP sulphurylase (selA) activities, while a further two classes (selD and selE) were pleiotropic and possibly regulatory. Most classes of sel mutations were clustered around the cysD locus of S. coelicolor. Segments of chromosomal DNA cloned from S. coelicolor, S. cattleya and S. clavuligerus and able to complement various sel/cys mutations allowed the relative positions of these mutations and the cysC and cysD mutations of S. coelicolor to be determined. The sel/cys DNA can be used for two-way selection: Cys+SelsCys-Selr. |
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Keywords: | Streptomyces Selenate resistance Cysteine auxotrophy Sulphate permease ATP sulphurylase |
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