Molecular cloning of the fnr gene of Escherichia coli K12

✍ Scribed by Shaw, Duncan J. ;Guest, John R.

• Publisher: Springer
• Year: 1981
• Tongue: English
• Weight: 594 KB
• Volume: 181
• Category: Article
• ISSN: 0026-8925
• DOI: 10.1007/bf00339011

✦ Synopsis

Mutations in the fnr gene of Escherichia coli have pleiotrophic effects leading to deficiencies in the reduction of fumarate and nitrate, hydrogen production and the ability to grow anaerobically with fumarate or nitrate as terminal electron acceptors. Transducing phages (lambda fnr) carrying the wild-type fnr gene were isolated from populations of artificially-constructed recombinant lambda phages by their ability to complement the lesions of fnr mutants. The lambda fnr phages restored anaerobic growth with fumarate and nitrate as electron acceptors and as prophages, they promoted normal synthesis of fumarate reductase, nitrate reductase and hydrogenase in fnr mutants. Five independently-isolated lambda fnr phages each contained a R.HindIII fragment (11.5 kilobases) that possessed three internal R.EcoRI targets and had inserted with the same orientation relative to the phage. A physical map of the fnr region was constructed by restriction analysis and flanking fragments were identified by DNA : DNA hybridization.