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Identification of essential genes and synthetic lethal gene combinations in Escherichia coli K-12. 2015
Hirotada Mori, and
Tomoya Baba, and
Katsushi Yokoyama, and
Rikiya Takeuchi, and
Wataru Nomura, and
Kazuichi Makishi, and
Yuta Otsuka, and
Hitomi Dose, and
Barry L Wanner
Graduate School of Biological Sciences, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma, Nara, 630-0101, Japan, hmori@gtc.naist.jp.
Here we describe the systematic identification of single genes and gene pairs, whose knockout causes lethality in Escherichia coli K-12. During construction of precise single-gene knockout library of E. coli K-12, we identified 328 essential gene candidates for growth in complex (LB) medium. Upon establishment of the Keio single-gene deletion library, we undertook the development of the ASKA single-gene deletion library carrying a different antibiotic resistance. In addition, we developed tools for identification of synthetic lethal gene combinations by systematic construction of double-gene knockout mutants. We introduce these methods herein.
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