产志贺毒素大肠杆菌中的 Lambda Red 介导的重组。
Lambda Red-Mediated Recombination in Shiga Toxin-Producing Escherichia coli.
机构信息
Department of Molecular & Cell Biology, Institute for Systems Genomics, University of Connecticut, Storrs, CT, USA.
出版信息
Methods Mol Biol. 2021;2291:145-162. doi: 10.1007/978-1-0716-1339-9_6.
The bacteriophage Lambda (λ) "Red" recombination system has enabled the development of efficient methods for engineering bacterial chromosomes. This system has been particularly important to the field of bacterial pathogenesis, where it has advanced the study of virulence factors from Shiga toxin-producing and enteropathogenic Escherichia coli (STEC and EPEC). Transient plasmid-driven expression of Lambda Red allows homologous recombination between PCR-derived linear DNA substrates and target loci in the STEC/EPEC chromosomes. Red-associated techniques can be used to create individual gene knockouts, generate deletions of large pathogenicity islands, and make markerless allelic exchanges. This chapter describes specific strategies and procedures for performing Lambda Red-mediated genome engineering in STEC.
噬菌体 Lambda（λ）“Red”重组系统为高效地构建细菌染色体工程提供了手段。该系统对细菌发病机制领域非常重要，特别是在志贺毒素产生大肠杆菌（STEC）和肠致病性大肠杆菌（EPEC）中，它促进了毒力因子的研究。Lambda Red 瞬时质粒驱动表达使 PCR 衍生的线性 DNA 底物与 STEC/EPEC 染色体中的靶标位点之间发生同源重组。与 Red 相关的技术可用于构建单个基因敲除、大致病性岛缺失和无标记等位基因交换。本章介绍了在 STEC 中进行 Lambda Red 介导的基因组工程的具体策略和程序。
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