High-resolution protein-protein interaction mapping using all-versus-all sequencing (AVA-Seq).

ABSTRACT: Two-hybrid systems can be used for investigating protein-protein interactions and may provide important information about gene products with unknown function. Despite their success in mapping protein interactions, two-hybrid systems have remained mostly untouched by improvements in next-generation DNA sequencing. The two-hybrid systems rely on one-versus-all methods in which each bait is sequentially screened against an entire library. Here, we developed a screening method that joins both bait and prey as a convergent fusion into one bacterial plasmid vector that can then be amplified and paired-end sequencing by next-generation sequencing (NGS). Our method enables all-versus-all sequencing (AVA-Seq) and utilizes NGS to remove multiple bottlenecks of the two-hybrid system. AV

SUBMITTER: Andrews SS 

PROVIDER: S-EPMC6663860 | biostudies-literature | 2019 Jul

REPOSITORIES: biostudies-literature