Moore Sean D
Burnett School of Biomedical Sciences, College of Medicine, University of Central Florida, Orlando, FL, USA.
Methods Mol Biol. 2011;765:155-69. doi: 10.1007/978-1-61779-197-0_10.
A protocol is described that allows the transfer of genetic material from one Escherichia coli strain to another using bacteriophage P1. P1 transduction can be used to construct new bacterial strains containing multiple alleles, to restore a locus to wild type, to move specific genetic markers from one strain to another, to relocate different mutant genes to a common genetic background, and to evaluate second-site suppression of a mutant allele. Because of these abilities, P1 transduction remains a staple in the arsenal of genetic tools that have kept E. coli at the forefront of model bacterial systems. The protocol incorporates some updated steps and discusses general principles of bacteriophage handling and the infection process.
本文描述了一种使用噬菌体P1将遗传物质从一种大肠杆菌菌株转移到另一种大肠杆菌菌株的方法。P1转导可用于构建含有多个等位基因的新细菌菌株、将一个位点恢复为野生型、将特定遗传标记从一个菌株转移到另一个菌株、将不同的突变基因重新定位到共同的遗传背景中,以及评估突变等位基因的第二位点抑制。由于这些能力,P1转导仍然是遗传工具库中的一个主要方法,这些遗传工具使大肠杆菌一直处于模式细菌系统的前沿。该方法纳入了一些更新步骤,并讨论了噬菌体处理和感染过程的一般原则。