Mitigating genetic instability caused by the excision activity of the <i>phi</i>C31 integrase in <i>Streptomyces</i>.

ABSTRACT: Over the past three decades, the integrase (Int) from Streptomyces phage phiC31 has become a valuable genome engineering tool across various species. phiC31 Int was thought to mediate unidirectional site-specific integration (attP × attB to attL and attR) in the absence of the phage-encoded recombination directionality factor (RDF). However, we have shown in this study that Int can also catalyze reverse excision (attL × attR to attP and attB) at low frequencies in Streptomyces lividans and Escherichia coli, causing genetic instability in engineered strains. To address this issue, we developed a two-plasmid co-conjugation (TPC) system. This system consists of an attP-containing integration vect

SUBMITTER: Duan Y 

PROVIDER: S-EPMC11784100 | biostudies-literature | 2025 Jan

REPOSITORIES: biostudies-literature