1887

Abstract

Recombineering using bacteriophage lambda Red recombinase (λ-Red) uses homologous recombination to manipulate bacterial genomes and is commonly applied to disrupt genes to elucidate their function. This is often followed by the introduction of a wild-type copy of the gene on a plasmid to complement its function. This is often not, however, at a native copy number and the introduction of a chromosomal version of a gene can be a desirable solution to provide wild-type copy expression levels of an allele . Here, we present a simple methodology based on the λ-Red-based ‘gene doctoring’ technique, where we developed tools used for chromosomal tagging in a conserved locus downstream of and found no impact on a variety of important phenotypes. The tools described provide an easy, quick and inexpensive method of chromosomal modification for the creation of a library of insertion mutants to study gene function.

Funding
This study was supported by the:
  • Biotechnology and Biological Sciences Research Council (Award BB/R012504/1)
    • Principle Award Recipient: Not Applicable
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/content/journal/micro/10.1099/mic.0.000994
2020-11-23
2021-07-26
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