A complete set of Escherichia coli open reading frames in mobile plasmids facilitating genetic studies.
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.
Saka
Kimiko
K
National Institute of Genetics, Mishima, Shizuoka 411-8540, Japan.
Tadenuma
Maki
M
Nakade
Shinsuke
S
Tanaka
Noriko
N
Sugawara
Hideaki
H
Nishikawa
Ken
K
Ichiyoshi
Nobuyuki
N
Kitagawa
Masanari
M
Mori
Hirotada
H
Ogasawara
Naotake
N
Nishimura
Akiko
A
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