Lambda Red Recombineering of Bacteriophage in the Lysogenic State

Matthew Tridgett; Maria Ababi; Alfonso Jaramillo

doi:10.1007/978-1-0716-2233-9_2

Lambda Red Recombineering of Bacteriophage in the Lysogenic State

Methods Mol Biol. 2022:2479:11-19. doi: 10.1007/978-1-0716-2233-9_2.

Authors

Matthew Tridgett¹, Maria Ababi^{1

2}, Alfonso Jaramillo^{3

4}

Affiliations

¹ School of Life Sciences, University of Warwick, Coventry, UK.
² Warwick Medical School, University of Warwick, Coventry, UK.
³ School of Life Sciences, University of Warwick, Coventry, UK. Alfonso.Jaramillo@synth-bio.org.
⁴ De novo Synthetic Biology Lab, I2SysBio, CSIC-University of Valencia, Paterna, Spain. Alfonso.Jaramillo@synth-bio.org.

PMID: 35583729
DOI: 10.1007/978-1-0716-2233-9_2

Abstract

We present a recombineering-based method for editing the genome of a temperate phage. The method uses the lambda Red recombination system to edit the genome of a lysogenized host with a prophage compatible with bacteriophage lambda. Linear DNA is used as the recombination substrate and antibiotic resistance is used as the basis for selection of recombinants. The method enables the genetic manipulation of a prophage in 3-5 days.

Keywords: Bacteriophage; Electroporation; Lambda red; Lysogenic; Recombineering; Temperate.

MeSH terms

Bacteriophage lambda / genetics
Escherichia coli* / genetics
Lysogeny / genetics
Prophages / genetics
Recombination, Genetic*