A complete set of Escherichia coli open reading frames in mobile plasmids facilitating genetic studies. |
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Authors: | Kimiko Saka Maki Tadenuma Shinsuke Nakade Noriko Tanaka Hideaki Sugawara Ken Nishikawa Nobuyuki Ichiyoshi Masanari Kitagawa Hirotada Mori Naotake Ogasawara Akiko Nishimura |
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Affiliation: | National Institute of Genetics, Mishima, Shizuoka 411-8540, Japan. |
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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. |
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Keywords: | Mobile plasmid clones ORF clone set cell division Escherichia coli |
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