LoxTnSeq: random transposon insertions combined with cre/lox recombination and counterselection to generate large random genome reductions.

ABSTRACT: The removal of unwanted genetic material is a key aspect in many synthetic biology efforts and often requires preliminary knowledge of which genomic regions are dispensable. Typically, these efforts are guided by transposon mutagenesis studies, coupled to deepsequencing (TnSeq) to identify insertion points and gene essentiality. However, epistatic interactions can cause unforeseen changes in essentiality after the deletion of a gene, leading to the redundancy of these essentiality maps. Here, we present LoxTnSeq, a new methodology to generate and catalogue libraries of genome reduction mutants. LoxTnSeq combines random integration of lox sites by transposon mutagenesis, and the generation of mutants via Cre recombinase, catalogued via deep sequencing. When LoxTnSeq was applied to the natur

SUBMITTER: Shaw D 

PROVIDER: S-EPMC8601177 | biostudies-literature | 2021 Nov

REPOSITORIES: biostudies-literature