A Yeast BiFC-seq Method for Genome-wide Interactome Mapping
- Author:
Shang LIMIN
1
;
Zhang YUEHUI
;
Liu YUCHEN
;
Jin CHAOZHI
;
Yuan YANZHI
;
Tian CHUNYAN
;
Ni MING
;
Bo XIAOCHEN
;
Zhang LI
;
Li DONG
;
He FUCHU
;
Wang JIAN
Author Information
1. State Key Laboratory of Proteomics,Beijing Proteome Research Center,National Center for Protein Sciences(Beijing),Beijing Institute of Lifeomics,Beijing 102206,China
- Keywords:
Bimolecular fluorescence complementation;
Protein-protein interaction;
High-throughput;
Next-generation sequencing
- From:
Genomics, Proteomics & Bioinformatics
2022;20(4):795-807
- CountryChina
- Language:Chinese
-
Abstract:
Genome-wide physical protein-protein interaction(PPI)mapping remains a major chal-lenge for current technologies.Here,we reported a high-efficiency BiFC-seq method,yeast-enhanced green fluorescent protein-based bimolecular fluorescence complementation(yEGFP-BiFC)coupled with next-generation DNA sequencing,for interactome mapping.We first applied yEGFP-BiFC method to systematically investigate an intraviral network of the Ebola virus.Two-thirds(9/14)of known interactions of EBOV were recaptured,and five novel interactions were discovered.Next,we used the BiFC-seq method to map the interactome of the tumor protein p53.We identified 97 interactors of p53,more than three-quarters of which were novel.Furthermore,in a more complex background,we screened potential interactors by pooling two BiFC libraries together and revealed a network of 229 interactions among 205 proteins.These results show that BiFC-seq is a highly sensitive,rapid,and economical method for genome-wide interactome map-ping.
