Genetic and physical mapping of an hydrogenase gene cluster from Rhodobacter capsulatus |
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Authors: | A. Colbeau J.-P. Magnin B. Cauvin T. Champion P. M. Vignais |
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Affiliation: | (1) Biochimie Microbienne/LBIO/DRF (CNRS UA1130), Centre d'Etudes Nucléaires de Grenoble, 85 X, F-38041 Grenoble Cedex, France;(2) Present address: CREMGP/ENSEEG Domaine Universitaire, BP 75, 38402 Saint-Martin-d'Hères Cedex, France |
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Abstract: | Summary An hydrogenase-deficient (Hup–) mutant of Rhodobacter capsulatus was obtained by adventitious insertion of IS21 DNA into an hydrogenase structural gene (hup) of the wild-type strain 1310. The resulting Hup– mutant, strain JP91, selected by its inability to grow autotrophically (Aut– phenotype) together with other Hup– mutant strains obtained by classical ethyl methane sulphonate mutagenesis were used in R plasmid-mediated conjugation experiments to map the hup/aut loci on the chromosome of R. capsulatus. The hup genes tested in this study were found to cluster on the chromosome in the proximity of the his-1 marker. A cluster of hup genes comprising the structural genes was isolated from a gene bank constructed in the cosmid vector pHC79 with 40 kb insert DNA. The clustered hup genes, characterized by hybridization studies and complementation analyses of the R. capsulatus Hup– mutants, span 15–20 kb of DNA. |
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Keywords: | Hydrogenase (hup) genes Rhodobacter capsulatus hupS, hupL (NiFe)hydrogenase IS21 Chromosomal mapping |
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