title: A genetic assay for gene essentiality in Clostridium.
abstract: Essential genes of pathogens are potential therapeutic targets, but are difficult to verify. Here, gene essentiality was determined by targeted knockout following engineered gene duplication. Null mutants of candidate essential genes of Clostridium difficile were viable only in the presence of a stable second copy of the gene.
authors.affiliation: Clostridia Research Group, BBSRC/EPSRC Synthetic Biology Research Centre (SBRC), School of Life Sciences, Centre for Biomolecular Sciences, University of Nottingham, University Park, Nottingham, NG7 2RD, United Kingdom.
authors.forename: David J F
authors.initials: DJF
authors.lastname: Walker
authors.affiliation: Clostridia Research Group, BBSRC/EPSRC Synthetic Biology Research Centre (SBRC), School of Life Sciences, Centre for Biomolecular Sciences, University of Nottingham, University Park, Nottingham, NG7 2RD, United Kingdom.
authors.forename: John T
authors.initials: JT
authors.lastname: Heap
authors.affiliation: Clostridia Research Group, BBSRC/EPSRC Synthetic Biology Research Centre (SBRC), School of Life Sciences, Centre for Biomolecular Sciences, University of Nottingham, University Park, Nottingham, NG7 2RD, United Kingdom.
authors.forename: Klaus
authors.initials: K
authors.lastname: Winzer
authors.affiliation: Clostridia Research Group, BBSRC/EPSRC Synthetic Biology Research Centre (SBRC), School of Life Sciences, Centre for Biomolecular Sciences, University of Nottingham, University Park, Nottingham, NG7 2RD, United Kingdom; Nottingham Digestive Disease Centre, NIHR Biomedical Research Unit, The University of Nottingham, University Park, Nottingham, United Kingdom. Electronic address: nigel.minton@nottingham.ac.uk.
authors.forename: Nigel P
authors.initials: NP
authors.lastname: Minton
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D001681:Biological Assay; D003001:Cloning, Molecular; D016360:Clostridioides difficile / Q000235:genetics*; D020440:Gene Duplication; D015870:Gene Expression; D020043:Genes, Essential; D005818:Genetic Engineering / Q000379:methods*; D008716:Methionine Adenosyltransferase / Q000235:genetics*; D014369:Tryptophan-tRNA Ligase / Q000235:genetics*
chemical_list: D008716:Methionine Adenosyltransferase; D014369:Tryptophan-tRNA Ligase
keywords: ClosTron; Clostridium difficile; Gene essentiality; Merodiploid; Therapeutic target