大肠杆菌K-12中必需基因和合成致死基因组合的鉴定

Identification of essential genes and synthetic lethal gene combinations in Escherichia coli K-12.

作者信息

Mori Hirotada, Baba Tomoya, Yokoyama Katsushi, Takeuchi Rikiya, Nomura Wataru, Makishi Kazuichi, Otsuka Yuta, Dose Hitomi, Wanner Barry L

机构信息

Graduate School of Biological Sciences, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma, Nara, 630-0101, Japan,

出版信息

Methods Mol Biol. 2015;1279:45-65. doi: 10.1007/978-1-4939-2398-4_4.

DOI:10.1007/978-1-4939-2398-4_4

PMID:25636612

Abstract

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.

摘要

在此，我们描述了对单个基因和基因对的系统鉴定，其敲除会导致大肠杆菌K-12致死。在构建大肠杆菌K-12精确单基因敲除文库的过程中，我们鉴定出328个在复杂（LB）培养基中生长所必需的候选基因。在建立了Keio单基因缺失文库后，我们着手开发携带不同抗生素抗性的ASKA单基因缺失文库。此外，我们通过系统构建双基因敲除突变体，开发了鉴定合成致死基因组合的工具。我们在此介绍这些方法。