Identification of Essential Genes and Synthetic Lethal Gene Combinations in Escherichia coli K-12
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.
Key wordsGene deletion Escherichia coli FLP recombinase FRT Gene knockout Homologous recombination In-frame deletion Lambda Red recombinase Essential genes KEIO collection ASKA collection
This work was supported by a Grant-in-Aid for Scientific Research on Priority Areas from the Ministry of Education, Culture, Sports, Science, and Technology of Japan and a grant from CREST, JST (Japan Science and Technology), NEDO (New Energy and Industrial Technology Development Organization), Tsuruoka City, Yamagata prefecture in Japan. B.L.W. is supported by the US National Science Foundation (grant number 106394).
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