Control of ϕC31 integrase-mediated site-specific recombination by protein trans-splicing.

ABSTRACT: Serine integrases are emerging as core tools in synthetic biology and have applications in biotechnology and genome engineering. We have designed a split-intein serine integrase-based system with potential for regulation of site-specific recombination events at the protein level in vivo. The ϕC31 integrase was split into two extein domains, and intein sequences (Npu DnaEN and Ssp DnaEC) were attached to the two termini to be fused. Expression of these two components followed by post-translational protein trans-splicing in Escherichia coli generated a fully functional ϕC31 integrase. We showed that protein splicing is necessary for recombination activity; deletion of intein domains or mutation of key intein residues inactivated recombination. We used an invertible promoter reporter system t

SUBMITTER: Olorunniji FJ 

PROVIDER: S-EPMC6868429 | biostudies-literature | 2019 Dec

REPOSITORIES: biostudies-literature