title: Attenuated colicin-based screening to discover and create novel resistance genes.
abstract: I formulated a systematic approach to screen for colicin resistance-associated genes by using an attenuated colicin mutant and demonstrated its utility in a screen of genes related to colicin E5 resistance. Screening of an Escherichia coli genome library revealed rstA as a partial resistance gene to colicin E5. Transcript expression of BtuB and OmpF, proteins responsible for translocation of nuclease E colicins, was clearly inhibited in an rstA-overexpressing strain. In addition, the tatA::recN fusion gene provides resistance to an attenuated colicin E5 mutant. Improving tatA::recN by directed coevolution, I created a novel gene with enhanced resistance to colicin E5 and concluded that attenuated colicin-based screening is useful for the discovery and creation of novel colicin resistance-associated genes.
authors.affiliation: Life Science Center of Tsukuba Advanced Research Alliance, University of Tsukuba, Japan. Electronic address: futaikazuki@gmail.com.
authors.forename: Kazuki
authors.initials: K
authors.lastname: Futai
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D003087:Colicins / Q000494:pharmacology*; D024881:Drug Resistance, Bacterial; D004926:Escherichia coli / Q000187:drug effects* / Q000235:genetics*; D005798:Genes, Bacterial; D005818:Genetic Engineering; D005820:Genetic Testing; D009154:Mutation; D012641:Selection, Genetic
chemical_list: D000900:Anti-Bacterial Agents; D003087:Colicins
keywords: Attenuated colicin mutant; Directed evolution; Fused gene; Resistance-associated gene screening; rstA