Efficient transposon mutagenesis mediated by an IPTG-controlled conditional suicide plasmid.

ABSTRACT:

Background

Transposon mutagenesis is highly valuable for bacterial genetic and genomic studies. The transposons are usually delivered into host cells through conjugation or electroporation of a suicide plasmid. However, many bacterial species cannot be efficiently conjugated or transformed for transposon saturation mutagenesis. For this reason, temperature-sensitive (ts) plasmids have also been developed for transposon mutagenesis, but prolonged incubation at high temperatures to induce ts plasmid loss can be harmful to the hosts and lead to enrichment of mutants with adaptive genetic changes. In addition, the ts phenotype of a plasmid is often strain- or species-specific, as it may become non-ts or suicidal in different bacterial species.

Results

We have engineered several

SUBMITTER: Naorem SS 

PROVIDER: S-EPMC6201506 | biostudies-literature | 2018 Oct

REPOSITORIES: biostudies-literature