Journal Article

A Complete Set of <i>Escherichia coli</i> Open Reading Frames in Mobile Plasmids Facilitating Genetic Studies

Kimiko Saka, Maki Tadenuma, Shinsuke Nakade, Noriko Tanaka, Hideaki Sugawara, Ken Nishikawa, Nobuyuki Ichiyoshi, Masanari Kitagawa, Hirotada Mori, Naotake Ogasawara and Akiko Nishimura

in DNA Research

Published on behalf of Kazusa DNA Research Institute

Volume 12, issue 1, pages 63-68
Published in print January 2005 | ISSN: 1340-2838
Published online January 2005 | e-ISSN: 1756-1663 | DOI: http://dx.doi.org/10.1093/dnares/12.1.63

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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/lacIq. 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 Fbacteria 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.

Keywords: Mobile plasmid clones; ORF clone set; cell division; Escherichia coli

Journal Article.  0 words. 

Subjects: Genetics and Genomics

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