Circular permutation profiling by deep sequencing libraries created using transposon mutagenesis.

ABSTRACT: Deep mutational scanning has been used to create high-resolution DNA sequence maps that illustrate the functional consequences of large numbers of point mutations. However, this approach has not yet been applied to libraries of genes created by random circular permutation, an engineering strategy that is used to create open reading frames that express proteins with altered contact order. We describe a new method, termed circular permutation profiling with DNA sequencing (CPP-seq), which combines a one-step transposon mutagenesis protocol for creating libraries with a functional selection, deep sequencing and computational analysis to obtain unbiased insight into a protein's tolerance to circular permutation. Application of this method to an adenylate kinase revealed that CPP-seq creates tw

SUBMITTER: Atkinson JT 

PROVIDER: S-EPMC6061844 | biostudies-literature | 2018 Jul

REPOSITORIES: biostudies-literature