title: A complete set of Escherichia coli open reading frames in mobile plasmids facilitating genetic studies.
abstract: To facilitate genetic studies of Escherichia coli, we constructed a complete set of mobile plasmid clones of intact open reading frames (ORFs). Their expression is strictly controlled by Ptac / lacI(q). The plasmids carrying each ORF were introduced into an F+ recA strain and stored in 96-well microtiter plates. In this way, 96 clones can be transferred simultaneously to F- bacteria using the conjugative system. This provides a convenient procedure for systematic identification of ORFs that suppress or complement mutations. We created two types of clone sets: the original set contained individual clones in 45 microtiter plates, and a second set contained pools of 48 clones stored in a single microtiter plate. Using these clone sets, we have identified 403 genes that can correct in trans the temperature-sensitive defect of cell division mutants, which would suggest multiple global regulators for bacterial cell division.
authors.affiliation: National Institute of Genetics, Mishima, Shizuoka 411-8540, Japan.
authors.forename: Kimiko
authors.initials: K
authors.lastname: Saka
authors.forename: Maki
authors.initials: M
authors.lastname: Tadenuma
authors.forename: Shinsuke
authors.initials: S
authors.lastname: Nakade
authors.forename: Noriko
authors.initials: N
authors.lastname: Tanaka
authors.forename: Hideaki
authors.initials: H
authors.lastname: Sugawara
authors.forename: Ken
authors.initials: K
authors.lastname: Nishikawa
authors.forename: Nobuyuki
authors.initials: N
authors.lastname: Ichiyoshi
authors.forename: Masanari
authors.initials: M
authors.lastname: Kitagawa
authors.forename: Hirotada
authors.initials: H
authors.lastname: Mori
authors.forename: Naotake
authors.initials: N
authors.lastname: Ogasawara
authors.forename: Akiko
authors.initials: A
authors.lastname: Nishimura
mesh_terms: D001483:Base Sequence; D002455:Cell Division / Q000235:genetics; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics*; D005798:Genes, Bacterial; D005816:Genetic Complementation Test / Q000379:methods; D008957:Models, Genetic; D008969:Molecular Sequence Data; D016366:Open Reading Frames / Q000235:genetics*; D010957:Plasmids