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Marker exchange mutagenesis of the hydN genes in Desulfovibrio fructosovorans
Author(s) -
Rousset M.,
Dermoun Z.,
Chippaux M.,
Bélaich J. P.
Publication year - 1991
Publication title -
molecular microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.857
H-Index - 247
eISSN - 1365-2958
pISSN - 0950-382X
DOI - 10.1111/j.1365-2958.1991.tb01922.x
Subject(s) - biology , mutagenesis , genetics , gene , computational biology , mutation
Summary A strain of Desulfovibrio fructosovorans deleted from the hydN [NiFe]hydrogenase structural gene was constructed. A plasmid carrying a 7 kb DNA fragment on which the hydN gene had been replaced by the npt reporter gene (kanamycin‐resistant, Kn R ) was introduced into D. fructosovorans by electroporation. Southern analysis of one Kn R clone demonstrated that the hydN gene had been eliminated by marker exchange. This mutant, which was devoid of the [NiFe]hydrogenase gene, still showed a 10% residual hydrogenase activity. Its ability to grow efficiently with H 2 as sole energy source is discussed. This is the first report, in a member of the sulphate‐reducing bacteria, of a successful transformation and concomitant homologous recombination leading to a fully controlled genotype.