Rapid engineering of bacterial reporter gene fusions by using Red recombination.

ABSTRACT: Reporter gene fusions are essential tools for the investigation of gene regulation. Such fusions are traditionally generated by transposon mutagenesis and identified by a suitable selection procedure. Alternatively, specific reporter fusions can be generated by cloning of DNA fragments containing promoters or other regulatory elements in reporter plasmids. Here, we describe a novel approach for the rapid generation of reporter gene fusions in single copies at defined positions in bacterial genomes. This technique utilizes the Red recombinase for the homologous recombination of PCR-generated cassettes containing various currently used reporter genes, such as those for beta-galactosidase, luciferase, and green fluorescent protein. The approach allows the generation of transcriptional or tran

SUBMITTER: Gerlach RG 

PROVIDER: S-EPMC1932792 | biostudies-literature | 2007 Jul

REPOSITORIES: biostudies-literature